Pub Date : 2009-12-01DOI: 10.1017/S1479236209990106
Wang Di, Lin Shaowu, Xu Gefeng, Liu Yang, Mou Zhenbo, Lu Tongyan
The genetic diversities of 72 individuals from three wild Lenok populations of Mudanjiang River (MD), Yalujiang River (YL) and Wusulijiang River (WSL) in the northeast of China were analysed using amplified fragment length polymorphism (AFLP) markers. The results showed that 541 polymorphic loci out of 559 were amplified by 12 primer pairs and the percentage of polymorphic loci was 96.78%. Shannon indices for the MD, YL and WSL populations were 0.3988±0.2913, 0.3254±0.3037, 0.2125±0.2862, respectively, and Nei's gene diversity indices were 0.2737±0.2062, 0.2229±0.2129, 0.1446±0.1985, respectively. The average total genetic diversity ( H t ) was 0.3512±0.0.0208 and the average genetic diversity within populations ( H s ) was 0.2137±0.0152. Among the three populations, the average genetic distance ( D st ) was 0.1375 and the gene differentiation coefficient ( G st ) was 0.3914. The genetic diversity was 60.85% within populations and 39.15% among populations. The gene flow index ( N m ) was 0.7776. The analysis of molecular variance (AMOVA) indicated that the average fixation index ( F st ) was 0.55336. The variance was 55.16% within populations and 44.84% among populations. The highest polymorphism ratio was in the MD group and the lowest in the WSL group.
{"title":"Assessing genetic diversity in three wild Brachymystax lenok populations using AFLP markers.","authors":"Wang Di, Lin Shaowu, Xu Gefeng, Liu Yang, Mou Zhenbo, Lu Tongyan","doi":"10.1017/S1479236209990106","DOIUrl":"https://doi.org/10.1017/S1479236209990106","url":null,"abstract":"The genetic diversities of 72 individuals from three wild Lenok populations of Mudanjiang River (MD), Yalujiang River (YL) and Wusulijiang River (WSL) in the northeast of China were analysed using amplified fragment length polymorphism (AFLP) markers. The results showed that 541 polymorphic loci out of 559 were amplified by 12 primer pairs and the percentage of polymorphic loci was 96.78%. Shannon indices for the MD, YL and WSL populations were 0.3988±0.2913, 0.3254±0.3037, 0.2125±0.2862, respectively, and Nei's gene diversity indices were 0.2737±0.2062, 0.2229±0.2129, 0.1446±0.1985, respectively. The average total genetic diversity ( H t ) was 0.3512±0.0.0208 and the average genetic diversity within populations ( H s ) was 0.2137±0.0152. Among the three populations, the average genetic distance ( D st ) was 0.1375 and the gene differentiation coefficient ( G st ) was 0.3914. The genetic diversity was 60.85% within populations and 39.15% among populations. The gene flow index ( N m ) was 0.7776. The analysis of molecular variance (AMOVA) indicated that the average fixation index ( F st ) was 0.55336. The variance was 55.16% within populations and 44.84% among populations. The highest polymorphism ratio was in the MD group and the lowest in the WSL group.","PeriodicalId":236932,"journal":{"name":"Chinese Journal of Agricultural Biotechnology","volume":"7 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2009-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"128549525","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2009-12-01DOI: 10.1017/S1479236209990064
Yang Li, Wang Xianzhong, Yang Meng-bo, Zhang Jia-hua
To illustrate the effect of nitric oxide (NO) on the microtubules of Sertoli cells (SC), SCs of piglets were treated with sodium nitroprusside (SNP). Changes in cell viability, anti-oxidant activity, enzyme activity and p38 mutagen-activated protein kinase (p38MAPK) activation were detected. The results were as follows. A low concentration of NO can keep SC microtubule and cell viability normal, and a high concentration of NO could increase p38MAPK activation, decrease anti-oxidant activity and transferrin secretion, and destroy the structure and distribution of the microtubules. The results suggest that SNP treatment results in an increase in NO in SCs and decreased cell anti-oxidant activity. The high concentration of NO destroys cell microtubules by activating p38MAPK.
{"title":"Effect of Nitric Oxide on Sertoli Cell Microtubule of Piglets","authors":"Yang Li, Wang Xianzhong, Yang Meng-bo, Zhang Jia-hua","doi":"10.1017/S1479236209990064","DOIUrl":"https://doi.org/10.1017/S1479236209990064","url":null,"abstract":"To illustrate the effect of nitric oxide (NO) on the microtubules of Sertoli cells (SC), SCs of piglets were treated with sodium nitroprusside (SNP). Changes in cell viability, anti-oxidant activity, enzyme activity and p38 mutagen-activated protein kinase (p38MAPK) activation were detected. The results were as follows. A low concentration of NO can keep SC microtubule and cell viability normal, and a high concentration of NO could increase p38MAPK activation, decrease anti-oxidant activity and transferrin secretion, and destroy the structure and distribution of the microtubules. The results suggest that SNP treatment results in an increase in NO in SCs and decreased cell anti-oxidant activity. The high concentration of NO destroys cell microtubules by activating p38MAPK.","PeriodicalId":236932,"journal":{"name":"Chinese Journal of Agricultural Biotechnology","volume":"22 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2009-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"133469656","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2009-12-01DOI: 10.1017/S1479236209990519
Wang Gong-jin, Zhou Xiao-long, Tan Xiao-dong, Yu Jianning, Xu Xiao-bo, F. Bi-qin
Mature porcine oocytes containing first polar bodies (Pb I) were obtained by in vitro culture of follicle oocytes from ovaries obtained from a local abattoir, and zygotes with second polar bodies (Pb II) were grown after in vitro fertilization of the mature oocytes. Extrusion, biological activity and morphology of Pb I and Pb II were statistically analysed. Polar bodies were isolated and collected from oocytes by enzyme digestion or micromanipulation. Their vigour under different preservation conditions was analysed and evaluated using a Trypan blue staining method. The results showed that 66.7% of the oocytes extruded Pb I after 40 h of in vitro mature culture of oocytes, and 49.7% of the zygotes extruded Pb II 20 h after in vitro fertilization. The efficiency of isolation of Pb II by micromanipulation significantly exceeded that by enzyme digestion, the Pb I and Pb II isolated by micromanipulation presenting with good vigour and normal morphology (95.3% versus 58.9%). The survival rates of Pb I and Pb II were 63.3% and 93.1% for 4 h at 39°C, 85.0% and 72.9% for 40 h at 4°C, and over 95.0% and 84.6% for less than 7 days at −20°C. In comparison with the above preservation conditions for Pb I and Pb II, the results for cryopreservation were best, with rates of survival as high as 89.1% for Pb I and 87.9% for Pb II for preservation periods of over a month, and rates of normal morphology of 97.8% and 95.7%, respectively. The Pb I and Pb II could be isolated and preserved effectively, for use in further research on the recombination of oocytes and zygotes.
{"title":"Collection and preservation of porcine polar bodies.","authors":"Wang Gong-jin, Zhou Xiao-long, Tan Xiao-dong, Yu Jianning, Xu Xiao-bo, F. Bi-qin","doi":"10.1017/S1479236209990519","DOIUrl":"https://doi.org/10.1017/S1479236209990519","url":null,"abstract":"Mature porcine oocytes containing first polar bodies (Pb I) were obtained by in vitro culture of follicle oocytes from ovaries obtained from a local abattoir, and zygotes with second polar bodies (Pb II) were grown after in vitro fertilization of the mature oocytes. Extrusion, biological activity and morphology of Pb I and Pb II were statistically analysed. Polar bodies were isolated and collected from oocytes by enzyme digestion or micromanipulation. Their vigour under different preservation conditions was analysed and evaluated using a Trypan blue staining method. The results showed that 66.7% of the oocytes extruded Pb I after 40 h of in vitro mature culture of oocytes, and 49.7% of the zygotes extruded Pb II 20 h after in vitro fertilization. The efficiency of isolation of Pb II by micromanipulation significantly exceeded that by enzyme digestion, the Pb I and Pb II isolated by micromanipulation presenting with good vigour and normal morphology (95.3% versus 58.9%). The survival rates of Pb I and Pb II were 63.3% and 93.1% for 4 h at 39°C, 85.0% and 72.9% for 40 h at 4°C, and over 95.0% and 84.6% for less than 7 days at −20°C. In comparison with the above preservation conditions for Pb I and Pb II, the results for cryopreservation were best, with rates of survival as high as 89.1% for Pb I and 87.9% for Pb II for preservation periods of over a month, and rates of normal morphology of 97.8% and 95.7%, respectively. The Pb I and Pb II could be isolated and preserved effectively, for use in further research on the recombination of oocytes and zygotes.","PeriodicalId":236932,"journal":{"name":"Chinese Journal of Agricultural Biotechnology","volume":"34 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2009-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"122577605","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2009-12-01DOI: 10.1017/S1479236209990441
L. Bin, Zhang Liping, W. Jianping, Ma Bin-Yun, Gao Feng-qin, Y. Lian
The domestic yak(Bos grunniens)metallothionein-Ⅰ(MT-Ⅰ)and metallothionein-Ⅲ(MT-Ⅲ)cDNA 3'-end full length sequences(331 and 378 bp,GenBank accession MT-ⅠNo.AY758557,MT-ⅢNo.DQ492300)from total RNA of liver and brain tissues were amplified and cloned by RT-PCR and 3'-RACE using the primers Y_(MT-Ⅰ)SP,Y_(MT-Ⅲ)SP and M13 primers M4, respectively,in which included MT-Ⅰ(183 bp)coding sequence,MT-Ⅲ(207 bp)cDNA coding sequence,and tailing signal AATAAA and Poly(A)at the 3'-end ofMT-Ⅰand MT-Ⅲ,respectively.The analyses showed that the yak cDNA sequence coding for MT-Ⅰprotein was composed of 61 amino acids,including 20 cysteines which had conserved tripetide structure,such as C-X-C,C-C-X-C-C, C-X-X-C,etc..The yak cDNA sequence coding for MT-Ⅲprotein was composed of 68 amino acids,including 19 cysteines which had both MT-Ⅲitself specific conserved tripetides and the same conserved tripetides with MT-Ⅰ,such as T,CPCP,GEGAEA,etc..They were all comparatively conservative in molecule evolution.These structures indicated that MT-Ⅲhad both the same physiological functions of heavy metals disintoxicating etc.with MT-Ⅰand the different specific functions,which inhibits the neurons growth,from MT-Ⅰ.
{"title":"Cloning of 3′-end full-length cDNA encoding for MT-I and MT-III in yak and analysis of their sequences.","authors":"L. Bin, Zhang Liping, W. Jianping, Ma Bin-Yun, Gao Feng-qin, Y. Lian","doi":"10.1017/S1479236209990441","DOIUrl":"https://doi.org/10.1017/S1479236209990441","url":null,"abstract":"The domestic yak(Bos grunniens)metallothionein-Ⅰ(MT-Ⅰ)and metallothionein-Ⅲ(MT-Ⅲ)cDNA 3'-end full length sequences(331 and 378 bp,GenBank accession MT-ⅠNo.AY758557,MT-ⅢNo.DQ492300)from total RNA of liver and brain tissues were amplified and cloned by RT-PCR and 3'-RACE using the primers Y_(MT-Ⅰ)SP,Y_(MT-Ⅲ)SP and M13 primers M4, respectively,in which included MT-Ⅰ(183 bp)coding sequence,MT-Ⅲ(207 bp)cDNA coding sequence,and tailing signal AATAAA and Poly(A)at the 3'-end ofMT-Ⅰand MT-Ⅲ,respectively.The analyses showed that the yak cDNA sequence coding for MT-Ⅰprotein was composed of 61 amino acids,including 20 cysteines which had conserved tripetide structure,such as C-X-C,C-C-X-C-C, C-X-X-C,etc..The yak cDNA sequence coding for MT-Ⅲprotein was composed of 68 amino acids,including 19 cysteines which had both MT-Ⅲitself specific conserved tripetides and the same conserved tripetides with MT-Ⅰ,such as T,CPCP,GEGAEA,etc..They were all comparatively conservative in molecule evolution.These structures indicated that MT-Ⅲhad both the same physiological functions of heavy metals disintoxicating etc.with MT-Ⅰand the different specific functions,which inhibits the neurons growth,from MT-Ⅰ.","PeriodicalId":236932,"journal":{"name":"Chinese Journal of Agricultural Biotechnology","volume":"1 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2009-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"128009547","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2009-12-01DOI: 10.1017/S1479236209990076
Lin Ya-qiu, Zhuang He-Lin, Yang Gongshe
By means of semi-quantitative reverse transcription-polymerase chain reaction (semi-qRT-PCR), cell transfection, Hoechst 33342 , AO and flow cytometry methods, the apoptotic change of porcine preadipocyte treated with 9-cis retinoic acid(9-cis- RA) of retinoid X receptor α(RXRα) ligand, transfected with pEnhanced green fluorescent proteins C2-RXRα (pEGFPC2-RXRα) and RXRα-small interfering RNA (RXRα-siRNA), was investigated respectively. The morphologic change of apoptosis in treated porcine preadipocyte was observed. It appeared the cell constriction, cellular shrinkage and condensation of chromatin, pyknosis of nucleus in- to compact body firstly, and then fragmentation with completely nucleus membrane, budding and shedding continuously, at last cell turns to several apoptotic bodies enwraped with membrane in different size. Moreover, compared with control group, apoptotic rate was significantly decreased in 10 nmol/L 9-cisRA treated group, and higher in the 10 μmol/L 9-cisRA treated group (P 0.05). The expression of RXRα mRNA was significantly upregulated and downregulated when transfected with pEGFPC2-RXRα and RXRα-siRNA(P 0.05), respectively, and the apoptotic rate was significantly decreased in pEGFPC2-RXRα group while RXRα-siR- NA group significantly higher compared with control group (P 0.05). It implies that RXRα inhibits the apoptosis of porcine preadipocyte.
{"title":"Effect of retinoid X receptor α (RXRα) on apoptosis in primary cultured porcine pre-adipocytes.","authors":"Lin Ya-qiu, Zhuang He-Lin, Yang Gongshe","doi":"10.1017/S1479236209990076","DOIUrl":"https://doi.org/10.1017/S1479236209990076","url":null,"abstract":"By means of semi-quantitative reverse transcription-polymerase chain reaction (semi-qRT-PCR), cell transfection, Hoechst 33342 , AO and flow cytometry methods, the apoptotic change of porcine preadipocyte treated with 9-cis retinoic acid(9-cis- RA) of retinoid X receptor α(RXRα) ligand, transfected with pEnhanced green fluorescent proteins C2-RXRα (pEGFPC2-RXRα) and RXRα-small interfering RNA (RXRα-siRNA), was investigated respectively. The morphologic change of apoptosis in treated porcine preadipocyte was observed. It appeared the cell constriction, cellular shrinkage and condensation of chromatin, pyknosis of nucleus in- to compact body firstly, and then fragmentation with completely nucleus membrane, budding and shedding continuously, at last cell turns to several apoptotic bodies enwraped with membrane in different size. Moreover, compared with control group, apoptotic rate was significantly decreased in 10 nmol/L 9-cisRA treated group, and higher in the 10 μmol/L 9-cisRA treated group (P 0.05). The expression of RXRα mRNA was significantly upregulated and downregulated when transfected with pEGFPC2-RXRα and RXRα-siRNA(P 0.05), respectively, and the apoptotic rate was significantly decreased in pEGFPC2-RXRα group while RXRα-siR- NA group significantly higher compared with control group (P 0.05). It implies that RXRα inhibits the apoptosis of porcine preadipocyte.","PeriodicalId":236932,"journal":{"name":"Chinese Journal of Agricultural Biotechnology","volume":"18 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2009-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"115511857","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2009-12-01DOI: 10.1017/S1479236209990520
S. Zhen-hui, Guo Wan-zhu, Zhang Ying-jun
The recombinant PET-N plasmid, which includes the N gene of Transmissible gastroenteritis virus (TGEV), was transformed into Escherichia coli BL21(DE3) and expressed by 1.0 mmol/L IPTG at 37 ℃. The indirect ELISA for detecting TGEV nucleocapsid protein antibody was established after the reactionogenicity of the recombinant protein was proved by Western blot. The optional working circumstances for the ELISA are as follows: antigen concentration 15 μg/mL, serum dilution 1∶40, blocking solution 0.5% FBS, serum sample incubated for 90 min, concentration of HRP-spa 1∶5 000 incubated for 60 min, the substrate incubated at room temperature for 10 min, and the threshold value of ELISA OD450≥0.35 tried out with chess titration. Sensitivity , specificity and concordance of this method were 93.5%, 93.8% and 93.5%, respectively compared to Svanova TGEV/PRCV antibody diagnosis kit.
{"title":"Prokaryotic expression of nucleoprotein gene of Transmissible gastroenteritis virus and development of ELISA based on the expressed protein.","authors":"S. Zhen-hui, Guo Wan-zhu, Zhang Ying-jun","doi":"10.1017/S1479236209990520","DOIUrl":"https://doi.org/10.1017/S1479236209990520","url":null,"abstract":"The recombinant PET-N plasmid, which includes the N gene of Transmissible gastroenteritis virus (TGEV), was transformed into Escherichia coli BL21(DE3) and expressed by 1.0 mmol/L IPTG at 37 ℃. The indirect ELISA for detecting TGEV nucleocapsid protein antibody was established after the reactionogenicity of the recombinant protein was proved by Western blot. The optional working circumstances for the ELISA are as follows: antigen concentration 15 μg/mL, serum dilution 1∶40, blocking solution 0.5% FBS, serum sample incubated for 90 min, concentration of HRP-spa 1∶5 000 incubated for 60 min, the substrate incubated at room temperature for 10 min, and the threshold value of ELISA OD450≥0.35 tried out with chess titration. Sensitivity , specificity and concordance of this method were 93.5%, 93.8% and 93.5%, respectively compared to Svanova TGEV/PRCV antibody diagnosis kit.","PeriodicalId":236932,"journal":{"name":"Chinese Journal of Agricultural Biotechnology","volume":"7 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2009-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"117235353","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2009-12-01DOI: 10.1017/S1479236209990155
Liu-di Yuan, Meng Guo-Quan, Z. Jianping, Z. Teng, F. Juan, Ren Zheng-long
According to the known partial cDNA sequence of gibberellin-induced cysteine-rich protein from Gymnadnig conopsea,the primers bearing restriction enzyme site of EcoR I and HindⅢ were designed for amplifying the full-length of ORF and the signal peptide-truncated fragment of gcgasa gene. Two fragments with the length of 319 and 238 bp were obtained and then cloned into the plasmid pET-32(a). Following the transformation into Escherichia coli BL21(DE3) , the fusion proteins were observed at approximately 26.0 and 25.2 kD with the induction of 1 mmol/L IPTG. The results of SDS-PAGE and transmission electron micrograph of ultrathin section revealed that the presence of signal peptide gave rise to the formation of inclusion body, which was located in periplasmic space, however, the absence of signal peptide greatly enhanced the solubility of the target protein, and the expressed soluble protein was further purified by Ni2+-NTA affinity chromatography and gel filtration methods.
{"title":"Expression, purification and identification of gibberellin-induced cysteine-rich protein of Gymanadenia conopsea","authors":"Liu-di Yuan, Meng Guo-Quan, Z. Jianping, Z. Teng, F. Juan, Ren Zheng-long","doi":"10.1017/S1479236209990155","DOIUrl":"https://doi.org/10.1017/S1479236209990155","url":null,"abstract":"According to the known partial cDNA sequence of gibberellin-induced cysteine-rich protein from Gymnadnig conopsea,the primers bearing restriction enzyme site of EcoR I and HindⅢ were designed for amplifying the full-length of ORF and the signal peptide-truncated fragment of gcgasa gene. Two fragments with the length of 319 and 238 bp were obtained and then cloned into the plasmid pET-32(a). Following the transformation into Escherichia coli BL21(DE3) , the fusion proteins were observed at approximately 26.0 and 25.2 kD with the induction of 1 mmol/L IPTG. The results of SDS-PAGE and transmission electron micrograph of ultrathin section revealed that the presence of signal peptide gave rise to the formation of inclusion body, which was located in periplasmic space, however, the absence of signal peptide greatly enhanced the solubility of the target protein, and the expressed soluble protein was further purified by Ni2+-NTA affinity chromatography and gel filtration methods.","PeriodicalId":236932,"journal":{"name":"Chinese Journal of Agricultural Biotechnology","volume":"34 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2009-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"125486405","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2009-12-01DOI: 10.1017/S1479236209990040
Miao Ming-Xing, Yuan Yu-guo, An Li-You, Zhao Jun-hui, Bai Ya-Jun, Guo Lei, C. Yong
To check the antibacterial activity, the recombinant human lactoferrin (rhLF) was extracted from the milk of transgenic mices (Mus musculus) by gel filtration chromatography. The rhLF from the milk of transgenic mices (PCL25 and AP) were analyzed by SDS-PAGE, Western blot and ELISA assay. The bacteriostatic properties of rhLF were tested by agar disc diffusion method. The results indicated that the concentration of the hLF in the milk of transgenic mice ranged from 7 to 8 mg/mL when judged by ELISA analysis, the recombinant protein expressed in the milk had the same molecular weight as the native protein (about 78 kD) and the rhLFs had a strong antibacterial activity on Escherichia coli and Salmonella.
{"title":"Purification and antibacterial activity of recombinant human lactoferrin in milk of transgenic mice.","authors":"Miao Ming-Xing, Yuan Yu-guo, An Li-You, Zhao Jun-hui, Bai Ya-Jun, Guo Lei, C. Yong","doi":"10.1017/S1479236209990040","DOIUrl":"https://doi.org/10.1017/S1479236209990040","url":null,"abstract":"To check the antibacterial activity, the recombinant human lactoferrin (rhLF) was extracted from the milk of transgenic mices (Mus musculus) by gel filtration chromatography. The rhLF from the milk of transgenic mices (PCL25 and AP) were analyzed by SDS-PAGE, Western blot and ELISA assay. The bacteriostatic properties of rhLF were tested by agar disc diffusion method. The results indicated that the concentration of the hLF in the milk of transgenic mice ranged from 7 to 8 mg/mL when judged by ELISA analysis, the recombinant protein expressed in the milk had the same molecular weight as the native protein (about 78 kD) and the rhLFs had a strong antibacterial activity on Escherichia coli and Salmonella.","PeriodicalId":236932,"journal":{"name":"Chinese Journal of Agricultural Biotechnology","volume":"22 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2009-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"132781314","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2009-12-01DOI: 10.1017/S1479236209990507
Shen Xiao-yan, Cong Guozheng, Chang Huiyun, Li Xiangtao, Xie Qing-ge
The potential relationship between the establishment of Foot-and-mouth disease virus (FMDV) persistent infection and gene variation was identified by investigating the variation of VP1 and 3ABC genes from yellow cattle persistent infection isolates. Five yellow cattle were inoculated on their tongue with 1.0×10 4 ID 50 /ml of FMDV O/Akesu/58 strain. After displaying clinical or subclinical signs, they probably became asymptomatic carriers. Oesophageal–pharyngeal fluids were collected monthly from the carriers with a probang and inoculated into a baby hamster kidney cell line (BHK-21); 12 FMDV isolates were obtained. The VP1 and 3ABC genes of the 12 isolates were then amplified by reverse-transcriptase polymerase chain reaction (RT-PCR). Cloning and sequencing revealed that the homology of the VP1 nucleotide and amino-acid sequence of all the isolates was above 98%, with no base deletion or insertion. When compared with the O/Akesu/58 FMDV strain, the homology of the VP1 nucleotide sequence of the isolates was only 85%, and that of the deduced amino-acid sequence only 90%.There were several nucleotide mutations in the VP1 gene of the isolates, including 16 consistent nucleotide mutations, with only two of them leading to a change in amino acid (I 56 →T, A 210 →E). Moreover, it was found that four nucleotide points and three amino-acid points had transversions among all isolates. The 3ABC gene had only 13 nucleotide transversions and five amino-acid mutations. It was presumed that persistent FMDV infection might have little connection with variation in the VP1 and 3ABC genes, and was probably related to other structural protein gene and key factors.
通过对黄牛持续感染分离株VP1和3ABC基因变异的研究，确定了口蹄疫病毒(FMDV)持续感染的建立与基因变异之间的潜在关系。用1.0×10 4 id50 /ml的FMDV O/Akesu/58株在5头黄牛舌上接种。出现临床或亚临床症状后，可能成为无症状携带者。用probang每月从携带者身上收集食管咽液，并接种于幼鼠肾细胞系(BHK-21);分离得到12株口蹄疫病毒。利用逆转录聚合酶链反应(RT-PCR)扩增12株菌株的VP1和3ABC基因。克隆和测序结果表明，所有分离株VP1核苷酸和氨基酸序列同源性均在98%以上，无碱基缺失和插入。与O/Akesu/58 FMDV菌株的VP1核苷酸序列同源性仅为85%，推导出的氨基酸序列同源性仅为90%。分离株VP1基因存在多个核苷酸突变，包括16个一致的核苷酸突变，其中只有2个突变导致氨基酸的变化(I 56→T, a 210→E)。此外，发现4个核苷酸点和3个氨基酸点在所有分离株中都发生了翻转。3ABC基因只有13个核苷酸反转和5个氨基酸突变。推测FMDV持续感染可能与VP1和3ABC基因变异关系不大，可能与其他结构蛋白基因及关键因素有关。
{"title":"Variation analysis of the VP1 and 3ABC genes from yellow cattle isolates persistently infected by Foot-and-mouth disease virus.","authors":"Shen Xiao-yan, Cong Guozheng, Chang Huiyun, Li Xiangtao, Xie Qing-ge","doi":"10.1017/S1479236209990507","DOIUrl":"https://doi.org/10.1017/S1479236209990507","url":null,"abstract":"The potential relationship between the establishment of Foot-and-mouth disease virus (FMDV) persistent infection and gene variation was identified by investigating the variation of VP1 and 3ABC genes from yellow cattle persistent infection isolates. Five yellow cattle were inoculated on their tongue with 1.0×10 4 ID 50 /ml of FMDV O/Akesu/58 strain. After displaying clinical or subclinical signs, they probably became asymptomatic carriers. Oesophageal–pharyngeal fluids were collected monthly from the carriers with a probang and inoculated into a baby hamster kidney cell line (BHK-21); 12 FMDV isolates were obtained. The VP1 and 3ABC genes of the 12 isolates were then amplified by reverse-transcriptase polymerase chain reaction (RT-PCR). Cloning and sequencing revealed that the homology of the VP1 nucleotide and amino-acid sequence of all the isolates was above 98%, with no base deletion or insertion. When compared with the O/Akesu/58 FMDV strain, the homology of the VP1 nucleotide sequence of the isolates was only 85%, and that of the deduced amino-acid sequence only 90%.There were several nucleotide mutations in the VP1 gene of the isolates, including 16 consistent nucleotide mutations, with only two of them leading to a change in amino acid (I 56 →T, A 210 →E). Moreover, it was found that four nucleotide points and three amino-acid points had transversions among all isolates. The 3ABC gene had only 13 nucleotide transversions and five amino-acid mutations. It was presumed that persistent FMDV infection might have little connection with variation in the VP1 and 3ABC genes, and was probably related to other structural protein gene and key factors.","PeriodicalId":236932,"journal":{"name":"Chinese Journal of Agricultural Biotechnology","volume":"18 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2009-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"116146786","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2009-12-01DOI: 10.1017/S1479236209990222
Lv Yao-ping, C. Jianming, Ye Jinyun, Huang Xu-xiong, Lao Shen-Ying, Shen Binqian, Yao Zi-liang, Guo Jianlin, Y. Li-ping
Seven isoenergic semi-purified test diets containing graded levels of protein ranging from 20% to 50% were formulated using fish meal and casein as the protein sources. Triplicate groups of Barbudes caldwell juveniles with initial body weight of 1.26± 0.02 g respectively were fed by the test diets for 8 weeks. The results indicated that there was no significant effect of dietary protein levels on the survival rate, relative weight of viscera and relative weight of liver of the juvenile fish (P 0.05). The weight gain and specific growth rate of the fish were increased with dietary protein level increasing from 20% to 35% (P 0.05), but not affected significantly as dietary protein level increased from 35% to 50% (P 0.05). Feed efficiencies were not significantly different when fish fed diets with protein levels from 30% to 50%, but significantly higher than those of the other two treatments when fish fed diets with protein levels of 20% and 25% (P 0.05). The protein efficiency ratio(PER) was negatively correlated to the diet protein level(x) (PER = 3.006-0.03251x, R =0.9366). There was no significant effect of dietary protein levels on carcass moisture, crude protein and ash of the juveniles (P 0.05). While carcass lipid was decreased with the increase of dietary protein level (x) (L =8.2169-0.0458x, R =0.8551). There was no significant variation of hepato-pancreas protease activity among tests (P 0.05). Intestine protease activity and hepato-pancreas amylase activity were increased to some extent, and then decreased with continued increasing dietary protein level. The intestine amylase activity (IAA) of the juvenile was negatively correlated to dietary protein level (x) (IAA = 84.625-0.9147x, R =0.8463). It was estimated that the suitable protein level for the B. caldwell juvenile was 34% of dry diet when the broken-line model was introduced to regress the relationship of weight gain of the juvenile and dietary protein level.
以鱼粉和酪蛋白为蛋白质来源，配制了7种蛋白质含量为20% ~ 50%的等能半纯化试验饲料。选取初始体重为1.26±0.02 g的考德威尔巴布氏幼鱼饲喂3个重复组，饲喂试验饲料8周。结果表明，饲料蛋白质水平对幼鱼的成活率、内脏相对重量和肝脏相对重量均无显著影响(p0.05)。饲料蛋白质水平在20% ~ 35%范围内增加了鱼的增重和特定生长率(P 0.05)，但在35% ~ 50%范围内对鱼的增重和特定生长率影响不显著(P 0.05)。蛋白质水平为30% ~ 50%时，饲料效率差异不显著，但蛋白质水平为20%和25%时，饲料效率显著高于其他2个处理(P 0.05)。蛋白质效率(PER)与饲粮蛋白质水平(x)呈负相关(PER = 3.006 ~ 0.03251x, R =0.9366)。饲粮蛋白质水平对幼鱼胴体水分、粗蛋白质和灰分均无显著影响(p0.05)。胴体脂质随饲粮蛋白质水平的升高而降低(L =8.2169 ~ 0.0458x, R =0.8551)。各组肝胰腺蛋白酶活性差异无统计学意义(p0.05)。随着饲粮蛋白质水平的继续升高，肠道蛋白酶活性和肝胰淀粉酶活性均有一定程度的升高，随后又有所下降。幼鱼肠道淀粉酶活性(IAA)与饲粮蛋白质水平呈负相关(IAA = 84.625-0.9147x, R =0.8463)。采用折线模型对饲料蛋白质水平与幼鱼增重的关系进行回归，估计幼鱼适宜的蛋白质水平为干饲料的34%。
{"title":"The effects of diet protein levels on the growth, body composition and digestive enzyme activities of the Barbodes caldwelli juvenile","authors":"Lv Yao-ping, C. Jianming, Ye Jinyun, Huang Xu-xiong, Lao Shen-Ying, Shen Binqian, Yao Zi-liang, Guo Jianlin, Y. Li-ping","doi":"10.1017/S1479236209990222","DOIUrl":"https://doi.org/10.1017/S1479236209990222","url":null,"abstract":"Seven isoenergic semi-purified test diets containing graded levels of protein ranging from 20% to 50% were formulated using fish meal and casein as the protein sources. Triplicate groups of Barbudes caldwell juveniles with initial body weight of 1.26± 0.02 g respectively were fed by the test diets for 8 weeks. The results indicated that there was no significant effect of dietary protein levels on the survival rate, relative weight of viscera and relative weight of liver of the juvenile fish (P 0.05). The weight gain and specific growth rate of the fish were increased with dietary protein level increasing from 20% to 35% (P 0.05), but not affected significantly as dietary protein level increased from 35% to 50% (P 0.05). Feed efficiencies were not significantly different when fish fed diets with protein levels from 30% to 50%, but significantly higher than those of the other two treatments when fish fed diets with protein levels of 20% and 25% (P 0.05). The protein efficiency ratio(PER) was negatively correlated to the diet protein level(x) (PER = 3.006-0.03251x, R =0.9366). There was no significant effect of dietary protein levels on carcass moisture, crude protein and ash of the juveniles (P 0.05). While carcass lipid was decreased with the increase of dietary protein level (x) (L =8.2169-0.0458x, R =0.8551). There was no significant variation of hepato-pancreas protease activity among tests (P 0.05). Intestine protease activity and hepato-pancreas amylase activity were increased to some extent, and then decreased with continued increasing dietary protein level. The intestine amylase activity (IAA) of the juvenile was negatively correlated to dietary protein level (x) (IAA = 84.625-0.9147x, R =0.8463). It was estimated that the suitable protein level for the B. caldwell juvenile was 34% of dry diet when the broken-line model was introduced to regress the relationship of weight gain of the juvenile and dietary protein level.","PeriodicalId":236932,"journal":{"name":"Chinese Journal of Agricultural Biotechnology","volume":"32 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2009-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"121635927","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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