Pub Date : 2008-12-01DOI: 10.1017/S1479236208002404
Zhang Yihua, Dou Zhong-ying, Shen Wen-zheng, Yang Chun-rong, Gao Zhimin
The population doubling number(70～80 times) of human fetal bone marrow mesenchymal stem cells(BMMSCs) is about two times more than that (30～40 times) of the adult BMMSCs, and their differentiation capacity is superior to that of their adult counterparts. In this study, BMMSCs were isolated from 2- to 3-month-old human abortuses by scissoring their long bones lengthwise, followed by rinsing and culturing whole marrow cells. Basic medium and serum concentration for BMMSCs culture were optimized and growth curves made, both with MTT(3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di-phenytetrazoliumromide) reduction assay. Isolated cells were identified with flow-cytometry and immunocytochemistry for their antigen markers. The biosafety of isolated cells was evaluated by karyotype analysis and tumor forming experiment. The results indicated that lengthwise scissoring of fetal long bones and rinsing of their marrow cells was practical and useful to obtain the BMMSCs from human abortuses at the age of 2～3 months. In this experiment, α-MEM(minimum essential medium alpha medium) +20% FCS(fetal cattle serum) was the best system for the BMMSCs in vitro. The third passage BMMSCs expressed Oct4, SSEA3 and SSEA4 beside the surface markers of their adult counterparts. The population doubling time of the BMMSCs of passage 6, 12 and 24 were 34,36 and 40 h, respectively. The cells in all the passages showed diploid karyotype and formed no tumor in the nude mice. The BMMSCs of human abortuses at the age of 2～3 months were proved to be biologically safe and ideal seed cells for researches on human tissue engineering and regeneration medicine.
人胎儿骨髓间充质干细胞(BMMSCs)的群体倍增数(70~80倍)是成人骨髓间充质干细胞(30~40倍)的2倍左右，分化能力优于成人骨髓间充质干细胞。在这项研究中，从2至3个月大的流产患者中，通过纵向剪断其长骨，然后冲洗和培养整个骨髓细胞，分离出骨髓间充质干细胞。采用MTT(3-(4,5)-二甲基噻吩偶氮(-z-y1)-3,5-二苯四氮唑胺)还原法，优化BMMSCs培养的基本培养基和血清浓度，并绘制生长曲线。用流式细胞术和免疫细胞化学对分离细胞进行抗原标记鉴定。通过核型分析和肿瘤形成实验评价分离细胞的生物安全性。结果表明，将胎儿长骨纵向剪断并冲洗其骨髓细胞可获得2~3月龄人流产骨髓间充质干细胞。在本实验中，α-MEM(minimum essential medium - α medium) +20%胎牛血清是体外培养BMMSCs的最佳体系。第三代BMMSCs表达Oct4、SSEA3和SSEA4，并在其成体对应物的表面标记旁表达。传代6、12和24的BMMSCs群体倍增时间分别为34、36和40 h。所有传代细胞的核型均为二倍体，未在裸鼠体内形成肿瘤。2~3月龄人流产骨髓间充质干细胞具有良好的生物学安全性，是开展人体组织工程和再生医学研究的理想种子细胞。
{"title":"Isolation and culture of bone marrow mesenchymal stem cells from human fetus and their biological properties","authors":"Zhang Yihua, Dou Zhong-ying, Shen Wen-zheng, Yang Chun-rong, Gao Zhimin","doi":"10.1017/S1479236208002404","DOIUrl":"https://doi.org/10.1017/S1479236208002404","url":null,"abstract":"The population doubling number(70～80 times) of human fetal bone marrow mesenchymal stem cells(BMMSCs) is about two times more than that (30～40 times) of the adult BMMSCs, and their differentiation capacity is superior to that of their adult counterparts. In this study, BMMSCs were isolated from 2- to 3-month-old human abortuses by scissoring their long bones lengthwise, followed by rinsing and culturing whole marrow cells. Basic medium and serum concentration for BMMSCs culture were optimized and growth curves made, both with MTT(3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di-phenytetrazoliumromide) reduction assay. Isolated cells were identified with flow-cytometry and immunocytochemistry for their antigen markers. The biosafety of isolated cells was evaluated by karyotype analysis and tumor forming experiment. The results indicated that lengthwise scissoring of fetal long bones and rinsing of their marrow cells was practical and useful to obtain the BMMSCs from human abortuses at the age of 2～3 months. In this experiment, α-MEM(minimum essential medium alpha medium) +20% FCS(fetal cattle serum) was the best system for the BMMSCs in vitro. The third passage BMMSCs expressed Oct4, SSEA3 and SSEA4 beside the surface markers of their adult counterparts. The population doubling time of the BMMSCs of passage 6, 12 and 24 were 34,36 and 40 h, respectively. The cells in all the passages showed diploid karyotype and formed no tumor in the nude mice. The BMMSCs of human abortuses at the age of 2～3 months were proved to be biologically safe and ideal seed cells for researches on human tissue engineering and regeneration medicine.","PeriodicalId":236932,"journal":{"name":"Chinese Journal of Agricultural Biotechnology","volume":"182 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2008-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"123005723","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 : 2008-12-01DOI: 10.1017/S1479236208002131
Wang Yan, Hou Jia-fa
Using reverse transcription polymerase chain reaction (RT-PCR) and gene splicing by overlap extension (SOE-PCR), the DNA sequence encoding the chicken's receptor activator of NF -κB ligand (RANKL) was amplified, and then cloned into the expressing plasmid pET32a(+) with His-tag, and was highly expressed in Echerichia coli. Then the purified protein was added in primary cultures of chicken osteopclasts to observe its bioactivity. The results showed that the size of PCR product was 1 200 bp which was consistent with the expected one, and the relative molecular weight of induced protein was 64 kD. Western blot indicated that the induced protein could react with anti-His antibody. It was also found that the induced protein could stimulate mature chicken osteoclasts to resorb bone.
{"title":"Cloning, expression and bioactivity of chicken receptor activator of NF-κB ligand ( chRANKL )","authors":"Wang Yan, Hou Jia-fa","doi":"10.1017/S1479236208002131","DOIUrl":"https://doi.org/10.1017/S1479236208002131","url":null,"abstract":"Using reverse transcription polymerase chain reaction (RT-PCR) and gene splicing by overlap extension (SOE-PCR), the DNA sequence encoding the chicken's receptor activator of NF -κB ligand (RANKL) was amplified, and then cloned into the expressing plasmid pET32a(+) with His-tag, and was highly expressed in Echerichia coli. Then the purified protein was added in primary cultures of chicken osteopclasts to observe its bioactivity. The results showed that the size of PCR product was 1 200 bp which was consistent with the expected one, and the relative molecular weight of induced protein was 64 kD. Western blot indicated that the induced protein could react with anti-His antibody. It was also found that the induced protein could stimulate mature chicken osteoclasts to resorb bone.","PeriodicalId":236932,"journal":{"name":"Chinese Journal of Agricultural Biotechnology","volume":"42 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2008-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"133688728","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 : 2008-12-01DOI: 10.1017/S1479236208002325
Cao Dongmei, Fan Xi-Ying, Wang Yunshan, Kang Lifang
Activation tagging plays an important role in plant genomics studies. In this paper, an activation tagging library containing 50 000 Basta-resistant lines was constructed by mediated transformation of the mutant bzr1-D of Aribidopsis thaliana . Among these transformants, 47 lines showed obvious phenotypes, including late flowering time, dwarf growth habit, changed leaf shape, longer leaf petiole, leaves lacking trichomes, sterility and no kink between stem/leaf. Some T-DNA flanking sequences were obtained by TAIL (thermal asymmetric interlaced) and nested PCR. Results showed that the mutants contained one to three T-DNA insertions. The insertions were distributed in the first, fourth and fifth chromosomes of the genome.
{"title":"Activation tagging library construction and mutant phenotype analysis","authors":"Cao Dongmei, Fan Xi-Ying, Wang Yunshan, Kang Lifang","doi":"10.1017/S1479236208002325","DOIUrl":"https://doi.org/10.1017/S1479236208002325","url":null,"abstract":"Activation tagging plays an important role in plant genomics studies. In this paper, an activation tagging library containing 50 000 Basta-resistant lines was constructed by mediated transformation of the mutant bzr1-D of Aribidopsis thaliana . Among these transformants, 47 lines showed obvious phenotypes, including late flowering time, dwarf growth habit, changed leaf shape, longer leaf petiole, leaves lacking trichomes, sterility and no kink between stem/leaf. Some T-DNA flanking sequences were obtained by TAIL (thermal asymmetric interlaced) and nested PCR. Results showed that the mutants contained one to three T-DNA insertions. The insertions were distributed in the first, fourth and fifth chromosomes of the genome.","PeriodicalId":236932,"journal":{"name":"Chinese Journal of Agricultural Biotechnology","volume":"13 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2008-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"124547163","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 : 2008-12-01DOI: 10.1017/S1479236208002301
Gao Jun-feng, Zhen Xiao-feng, Ge Li-jun, L. Qing, R. Rong
An optimal protocol for cryopreservation of boar semen was established. First, the boar semen was pre-diluted with ZORLESCO (ZO) solution and pre-equilibrated at room temperature for 1 h. After adding extender I, spermatozoa were equilibrated at 5°C for 1.5 h; then an equal volume of extender II was added and the spermatozoa equilibrated for 2 h. The resulting spermatozoa were loaded into 0.25 ml straws, equilibrated for 10 min at 3 cm above the surface of liquid nitrogen (LN), then promptly submerged into LN. When thawing, straws were incubated in a water bath at 37°C for 30 s. This procedure yielded the highest post-thaw motility of 0.58±0.03 and plasma integrity of 63.2±1.2%, together with a normal acrosome in 51.4±2.6% of spermatozoa. Abnormal spermatozoa after freezing represented only 14.0±3.0%.
{"title":"Cryopreservation of boar semen using straws","authors":"Gao Jun-feng, Zhen Xiao-feng, Ge Li-jun, L. Qing, R. Rong","doi":"10.1017/S1479236208002301","DOIUrl":"https://doi.org/10.1017/S1479236208002301","url":null,"abstract":"An optimal protocol for cryopreservation of boar semen was established. First, the boar semen was pre-diluted with ZORLESCO (ZO) solution and pre-equilibrated at room temperature for 1 h. After adding extender I, spermatozoa were equilibrated at 5°C for 1.5 h; then an equal volume of extender II was added and the spermatozoa equilibrated for 2 h. The resulting spermatozoa were loaded into 0.25 ml straws, equilibrated for 10 min at 3 cm above the surface of liquid nitrogen (LN), then promptly submerged into LN. When thawing, straws were incubated in a water bath at 37°C for 30 s. This procedure yielded the highest post-thaw motility of 0.58±0.03 and plasma integrity of 63.2±1.2%, together with a normal acrosome in 51.4±2.6% of spermatozoa. Abnormal spermatozoa after freezing represented only 14.0±3.0%.","PeriodicalId":236932,"journal":{"name":"Chinese Journal of Agricultural Biotechnology","volume":"33 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2008-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"124307572","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 : 2008-12-01DOI: 10.1017/S147923620800243X
Li Ju-fen, M. Guo-bin, Xu Ling
The hybrid purity of melon ( Cucumis melo L.) was tested by polymerase chain reaction (PCR) assay based on simple sequence repeat (SSR) markers in two F 1 melon hybrids (‘Dongfangmi 1’ and ‘Dongfangmi 2’) and their parental lines. Twelve pairs of SSR primers for ‘Dongfangmi 1’ and three pairs for ‘Dongfangmi 2’ were selected. Results showed that self-inbred seeds were effectively distinguished from F 1 hybrid seeds using these SSR primers, a finding that was consistent with the results recorded from field tests. ‘Dongfangmi 1’ and ‘Dongfangmi 2’ were identified from their parental lines, and seven other uterine hybrid lines by multiplex primers MS48+MS60 and MS4+MS20, respectively. Contamination of F 1 hybrid seeds caused by self-inbred and other unknown pollens can be effectively and more reliably detected by PCR assays with multiplex SSR primers.
利用SSR标记对甜瓜(Cucumis melo L.) 2个f1甜瓜杂交种‘东方密1号’和‘东方密2号’及其亲本进行了杂种纯度检测。筛选出‘东方密1号’的12对SSR引物和‘东方密2号’的3对SSR引物。结果表明，利用这些SSR引物可以有效区分自交种子和f1杂交种种子，这与田间试验结果一致。利用多重引物MS48+MS60和MS4+MS20分别从亲本系和其他7个子宫杂交系中鉴定出“东方蜜1号”和“东方蜜2号”。利用多重SSR引物进行PCR检测，可以有效、可靠地检测自交花粉和其他未知花粉对f1杂交种种子的污染。
{"title":"SSR markers for identification of purity of melon hybrids","authors":"Li Ju-fen, M. Guo-bin, Xu Ling","doi":"10.1017/S147923620800243X","DOIUrl":"https://doi.org/10.1017/S147923620800243X","url":null,"abstract":"The hybrid purity of melon ( Cucumis melo L.) was tested by polymerase chain reaction (PCR) assay based on simple sequence repeat (SSR) markers in two F 1 melon hybrids (‘Dongfangmi 1’ and ‘Dongfangmi 2’) and their parental lines. Twelve pairs of SSR primers for ‘Dongfangmi 1’ and three pairs for ‘Dongfangmi 2’ were selected. Results showed that self-inbred seeds were effectively distinguished from F 1 hybrid seeds using these SSR primers, a finding that was consistent with the results recorded from field tests. ‘Dongfangmi 1’ and ‘Dongfangmi 2’ were identified from their parental lines, and seven other uterine hybrid lines by multiplex primers MS48+MS60 and MS4+MS20, respectively. Contamination of F 1 hybrid seeds caused by self-inbred and other unknown pollens can be effectively and more reliably detected by PCR assays with multiplex SSR primers.","PeriodicalId":236932,"journal":{"name":"Chinese Journal of Agricultural Biotechnology","volume":"1 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2008-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"130067325","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 : 2008-12-01DOI: 10.1017/S147923620800226X
Gao Fang-fang, Wu Zhongyi, Zeng Shen-ming
{"title":"Bovine interferon-tau expression in Escherichia coli and identification of its biological activities","authors":"Gao Fang-fang, Wu Zhongyi, Zeng Shen-ming","doi":"10.1017/S147923620800226X","DOIUrl":"https://doi.org/10.1017/S147923620800226X","url":null,"abstract":"","PeriodicalId":236932,"journal":{"name":"Chinese Journal of Agricultural Biotechnology","volume":"22 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2008-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"129171225","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 : 2008-12-01DOI: 10.1017/S1479236208002453
Song Ping, Wang Qin-ying, Wu Hui-xian, Lu Xiu-jun, W. Yong
{"title":"Identification of the cry gene in Bacillus thuringiensis strain WZ-9 and its toxicity against Henosepilachna vigintioctomaculata","authors":"Song Ping, Wang Qin-ying, Wu Hui-xian, Lu Xiu-jun, W. Yong","doi":"10.1017/S1479236208002453","DOIUrl":"https://doi.org/10.1017/S1479236208002453","url":null,"abstract":"","PeriodicalId":236932,"journal":{"name":"Chinese Journal of Agricultural Biotechnology","volume":"123 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2008-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"124155026","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 : 2008-12-01DOI: 10.1017/S1479236208002428
C. Xiaoyong, Tian Shujun, Sang Run-zi, Sun Shu-chun, Zhao Zhu-jun, L. Junjie, Dun Weitao
Effect of lamb age, transport stimulation and repeated hormone superovulation on the number of collected oocytes were determined, and effect of the culture medium containing ethylene-diamine-tetra-acetic acid (EDTA) on the development of produced embryos from lamb oocytes in vitro were studied. Results indicated that the mean number of collected and available oocytes from per 6- to 8- week lamb were 60.8±13.9 and 58.2±12.3 respectively, which were more than that those (27.3±5.1 and 26.0±4.9 ) from per 12- to 14- week lamb(P 0.05), respectively; The transport stimulation didn't decreased the number of collected oocytes from the superovulation lambs(P 0.05), however, the number of collected oocytes in the repeated superovulation group was reduced significantly compared with the control group(P 0.05); The embryonic culture medium containing 10 μmol/L EDTA could improve significantly the development capability of in vitro produced embryo from lamb oocytes (P 0.05), and the healthy lambs were born by the embryo transfer.
{"title":"Induction of lamb follicular development and embryo production in vitro","authors":"C. Xiaoyong, Tian Shujun, Sang Run-zi, Sun Shu-chun, Zhao Zhu-jun, L. Junjie, Dun Weitao","doi":"10.1017/S1479236208002428","DOIUrl":"https://doi.org/10.1017/S1479236208002428","url":null,"abstract":"Effect of lamb age, transport stimulation and repeated hormone superovulation on the number of collected oocytes were determined, and effect of the culture medium containing ethylene-diamine-tetra-acetic acid (EDTA) on the development of produced embryos from lamb oocytes in vitro were studied. Results indicated that the mean number of collected and available oocytes from per 6- to 8- week lamb were 60.8±13.9 and 58.2±12.3 respectively, which were more than that those (27.3±5.1 and 26.0±4.9 ) from per 12- to 14- week lamb(P 0.05), respectively; The transport stimulation didn't decreased the number of collected oocytes from the superovulation lambs(P 0.05), however, the number of collected oocytes in the repeated superovulation group was reduced significantly compared with the control group(P 0.05); The embryonic culture medium containing 10 μmol/L EDTA could improve significantly the development capability of in vitro produced embryo from lamb oocytes (P 0.05), and the healthy lambs were born by the embryo transfer.","PeriodicalId":236932,"journal":{"name":"Chinese Journal of Agricultural Biotechnology","volume":"221 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2008-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"131515225","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 : 2008-12-01DOI: 10.1017/S1479236208002313
Ren Xiao-hui, Lu Hu-ying, Liu Song-cai, Z. Mingjun, O. Song-ying, Li Hong-Yi, Zhang Yong-liang
The elements for Semliki forest virus (SFV) RNA replicon, a kind of new generation vector, comes from the Alphavirus genus. It was designed to overcome the poor efficacy of some current plasmid vector. Genes coding for viral replicases are reserved while genes coding for structure proteins are replaced by foreign gene in RNA replicon. High level replication of RNA and expression of foreign gene in cytoplasm are regulated by the replicases. To evaluate the effects of the SFV RNA replicon on the efficiency of gene expression, LacZ gene was inserted into pIRES-neo which digested by BamHⅠand dephosphorylated by shrimp alkaline phosphatase, and pIRES-neo-LacZ vector was constructed. RNA replicon vector pCMV-rep-LacZ and two conventional CMV promoter-based vector (pLNCX-LacZ and pIRES-neo-LacZ) were transfected by Lipofectin to prepared 293 cells, respectively. RNA replicon vector pCMV-Rep-GHRH(Growth hormone releasing hormone) and two conventional CMV promoter-based vector(pCDNA3.1(+)-GHRH and pIRES-neo-GHRH) were transfected by Lipofectin to prepared 293 cells, respectively, too. Results of quantitating for β-galactosidase expressed from transfected cells and quantitating for GHRH expressed from transfected cells by RIA and RT-PCR showed that the express level of RNA replicon vector was superior to ordinary vector and 2~3 times higher than normal plasmid vector. This will provide a beneficial exploring to improve the efficiency of gene expression in eukaryotic cell.
{"title":"Comparison of Conventional Plasmid Vector and Semliki forest virus-derived Vectors in Expressing Growth Hormone Releasing Hormone (GHRH)","authors":"Ren Xiao-hui, Lu Hu-ying, Liu Song-cai, Z. Mingjun, O. Song-ying, Li Hong-Yi, Zhang Yong-liang","doi":"10.1017/S1479236208002313","DOIUrl":"https://doi.org/10.1017/S1479236208002313","url":null,"abstract":"The elements for Semliki forest virus (SFV) RNA replicon, a kind of new generation vector, comes from the Alphavirus genus. It was designed to overcome the poor efficacy of some current plasmid vector. Genes coding for viral replicases are reserved while genes coding for structure proteins are replaced by foreign gene in RNA replicon. High level replication of RNA and expression of foreign gene in cytoplasm are regulated by the replicases. To evaluate the effects of the SFV RNA replicon on the efficiency of gene expression, LacZ gene was inserted into pIRES-neo which digested by BamHⅠand dephosphorylated by shrimp alkaline phosphatase, and pIRES-neo-LacZ vector was constructed. RNA replicon vector pCMV-rep-LacZ and two conventional CMV promoter-based vector (pLNCX-LacZ and pIRES-neo-LacZ) were transfected by Lipofectin to prepared 293 cells, respectively. RNA replicon vector pCMV-Rep-GHRH(Growth hormone releasing hormone) and two conventional CMV promoter-based vector(pCDNA3.1(+)-GHRH and pIRES-neo-GHRH) were transfected by Lipofectin to prepared 293 cells, respectively, too. Results of quantitating for β-galactosidase expressed from transfected cells and quantitating for GHRH expressed from transfected cells by RIA and RT-PCR showed that the express level of RNA replicon vector was superior to ordinary vector and 2~3 times higher than normal plasmid vector. This will provide a beneficial exploring to improve the efficiency of gene expression in eukaryotic cell.","PeriodicalId":236932,"journal":{"name":"Chinese Journal of Agricultural Biotechnology","volume":"104 ","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2008-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"120939593","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 : 2008-12-01DOI: 10.1017/S1479236208002416
Yu Fei, Geng Jian-hui, Ni Li-gang, He Xian-hong, Xu Qi, Li Bichun
{"title":"In vitro differentiation of chicken spermatogonial stem cells into adipocytes","authors":"Yu Fei, Geng Jian-hui, Ni Li-gang, He Xian-hong, Xu Qi, Li Bichun","doi":"10.1017/S1479236208002416","DOIUrl":"https://doi.org/10.1017/S1479236208002416","url":null,"abstract":"","PeriodicalId":236932,"journal":{"name":"Chinese Journal of Agricultural Biotechnology","volume":"14 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2008-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"131719169","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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