Pub Date : 2009-08-01DOI: 10.1017/S1479236209990301
Li Benjin, Lv Xin, Chen Qinghe, Lan Cheng-zhong, Zhao Jian, Qiu Rongzhou, Weng Qiyong
Simple sequence repeats (SSR) and random amplification of polymorphic DNA (RAPD) molecular markers were used to assess the genetic diversity of 80 isolates of Phytophthora infestans in potato (Solanum tuberosum) from Fujian, Heilongjiang, Hebei and Inner Mongolia Provinces in China. Polymorphism was identified by 13 SSR primers and 14 RAPD primers in the isolates of P. infestans in potato. A total of 76 bands were amplified by SSRs, with the percentage of polymorphic bands (PPB) being 78.9% and the similarity coefficient ranging between 0.00 and 0.42. A total of 189 bands were amplified by RAPDs, with the percentage of polymorphic bands being 95.2% and the similarity coefficient ranging between 0.04 and 0.66. Analysis of genetic diversity showed that there exists higher genetic variation in the Fujian population in comparison to the populations of Heilongjiang, Hebei and Inner Mongolia. Nei's genetic identity analysis indicates that the genetic similarity between populations of Heilongjiang and Inner Mongolia is the highest and that between Fujian and Hebei is the lowest. A cluster analysis revealed that isolates from Fujian, in the south of China, are distantly related to those from Heilongjiang, Hebei and Inner Mongolia in the north, and the Fujian population is distributed among more groups than the other three, exhibiting a higher genetic diversity.
{"title":"Population genetic diversity of Phytophthora infestans from China as revealed by SSRs and RAPDs","authors":"Li Benjin, Lv Xin, Chen Qinghe, Lan Cheng-zhong, Zhao Jian, Qiu Rongzhou, Weng Qiyong","doi":"10.1017/S1479236209990301","DOIUrl":"https://doi.org/10.1017/S1479236209990301","url":null,"abstract":"Simple sequence repeats (SSR) and random amplification of polymorphic DNA (RAPD) molecular markers were used to assess the genetic diversity of 80 isolates of Phytophthora infestans in potato (Solanum tuberosum) from Fujian, Heilongjiang, Hebei and Inner Mongolia Provinces in China. Polymorphism was identified by 13 SSR primers and 14 RAPD primers in the isolates of P. infestans in potato. A total of 76 bands were amplified by SSRs, with the percentage of polymorphic bands (PPB) being 78.9% and the similarity coefficient ranging between 0.00 and 0.42. A total of 189 bands were amplified by RAPDs, with the percentage of polymorphic bands being 95.2% and the similarity coefficient ranging between 0.04 and 0.66. Analysis of genetic diversity showed that there exists higher genetic variation in the Fujian population in comparison to the populations of Heilongjiang, Hebei and Inner Mongolia. Nei's genetic identity analysis indicates that the genetic similarity between populations of Heilongjiang and Inner Mongolia is the highest and that between Fujian and Hebei is the lowest. A cluster analysis revealed that isolates from Fujian, in the south of China, are distantly related to those from Heilongjiang, Hebei and Inner Mongolia in the north, and the Fujian population is distributed among more groups than the other three, exhibiting a higher genetic diversity.","PeriodicalId":236932,"journal":{"name":"Chinese Journal of Agricultural Biotechnology","volume":"41 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2009-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"125572827","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 : 2009-08-01DOI: 10.1017/S1479236209990192
J. Qin, L. Hua, Z. Hong, Wang Yu-ping, Liping Qiang, M. Daifu, X. Yiping, Li Qingchang
Amplified fragment length polymorphism (AFLP) markers linked to the stem nematode resistance gene were developed in sweet potato ( Ipomoea batatas (L.) Lam.). Using bulked segregant analysis (BSA), 800 AFLP primer combinations were screened in the resistant and susceptible bulked DNA from the 186 progeny of an F 1 single-cross population of Xu781 (resistant parent)×Xushu18 (susceptible parent), and 245 of these AFLP primers showed polymorphic bands between resistant and susceptible DNA. Primer combinations detecting polymorphism between the two bulks were used to screen the parents and eight individuals from each of the bulks. The results showed that E2M23 and E33M20 produced a specific band of about 500 bp and 200 bp in length, respectively, in the resistant plants but not in the susceptible plants, suggesting that the markers named E2M23 500 and E33M20 200 linked to a gene for stem nematode resistance. Amplified analysis of the 186 F 1 individuals indicated that the genetic distance between these two markers and the stem nematode resistance gene was 6.9 cM and 11.1 cM, respectively, measured with Mapmaker 3.0. These two AFLP markers were used to identify ten sweet potato varieties planted widely in China and the results were consistent with those of conventional resistance identification, indicating that the two markers can be used in molecular marker-assisted breeding for stem nematode resistance in the sweet potato.
{"title":"Development of AFLP markers linked to stem nematode resistance gene in sweet potato ( Ipomoea batatas (L.) Lam.)","authors":"J. Qin, L. Hua, Z. Hong, Wang Yu-ping, Liping Qiang, M. Daifu, X. Yiping, Li Qingchang","doi":"10.1017/S1479236209990192","DOIUrl":"https://doi.org/10.1017/S1479236209990192","url":null,"abstract":"Amplified fragment length polymorphism (AFLP) markers linked to the stem nematode resistance gene were developed in sweet potato ( Ipomoea batatas (L.) Lam.). Using bulked segregant analysis (BSA), 800 AFLP primer combinations were screened in the resistant and susceptible bulked DNA from the 186 progeny of an F 1 single-cross population of Xu781 (resistant parent)×Xushu18 (susceptible parent), and 245 of these AFLP primers showed polymorphic bands between resistant and susceptible DNA. Primer combinations detecting polymorphism between the two bulks were used to screen the parents and eight individuals from each of the bulks. The results showed that E2M23 and E33M20 produced a specific band of about 500 bp and 200 bp in length, respectively, in the resistant plants but not in the susceptible plants, suggesting that the markers named E2M23 500 and E33M20 200 linked to a gene for stem nematode resistance. Amplified analysis of the 186 F 1 individuals indicated that the genetic distance between these two markers and the stem nematode resistance gene was 6.9 cM and 11.1 cM, respectively, measured with Mapmaker 3.0. These two AFLP markers were used to identify ten sweet potato varieties planted widely in China and the results were consistent with those of conventional resistance identification, indicating that the two markers can be used in molecular marker-assisted breeding for stem nematode resistance in the sweet potato.","PeriodicalId":236932,"journal":{"name":"Chinese Journal of Agricultural Biotechnology","volume":"117 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2009-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"117281374","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 : 2009-08-01DOI: 10.1017/S1479236209990246
Jin Liang, L. Yun, Sun Zhen-yuan, L. Tianhong
{"title":"Cloning of carnation GA 20-oxidase and the construction of a plant RNAi vector","authors":"Jin Liang, L. Yun, Sun Zhen-yuan, L. Tianhong","doi":"10.1017/S1479236209990246","DOIUrl":"https://doi.org/10.1017/S1479236209990246","url":null,"abstract":"","PeriodicalId":236932,"journal":{"name":"Chinese Journal of Agricultural Biotechnology","volume":"73 3 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2009-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"128102079","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 : 2009-08-01DOI: 10.1017/S1479236209002654
Y. Ying, Lin Xiao-hui, Kong Xiang-mei, Jia Zhi-lei, Lin Chun-Jing, L. Cong-cong, Liu Na, Sun Chuan-bo, Li De-pu
Six transgenic maize lines with the fully modified gene GFM Cry1A were obtained and polymerase chain reaction (PCR) analysis of their T 5 generation plants indicated that foreign genes could be stably inherited. Three hybrids (Simi25 Bt , Tongdan24 Bt and Jidan209 Bt ) were created using the transgenic lines and combined with regular maize inbred lines. The results of resistance identification of the transgenic inbred lines and hybrids showed that the difference of resistance among transgenic lines was very significant; there also existed a difference among individuals of the same line and among the three hybrids. Compared with the control inbred line (CK), the average of four transgenic inbred lines in the number of tunnels per stalk, larval tunnel length per plant and length of surviving larvae were decreased by 70%, 80% and 70%, respectively. Enzyme-linked immunosorbent assay (ELISA) testing showed that the average of Bt Cry1A protein expression level for six transgenic inbred lines was 0.16% total protein and the highest expression level was 0.19%. At maturity, compared with the control variety (CK), the larval tunnel length per plant of the three hybrids (Simi25 Bt , Tongdan24 Bt and Jidan209 Bt ) was decreased by 39.97%, 36.20% and 53.83%, respectively, which was a decrease of 43.36% on average. The investigation of agronomic characters showed that there was no significant difference between the improved hybrids and the control in plant height, ear length, row number per ear, kernel number per row and 100-kernels weight. It is thought that the GFM Cry1A gene can be applied to improve resistance to corn-borer, and maize inbred lines with the Bt gene can be directly applied to conventional maize breeding.
{"title":"Identification of resistance to corn-borer of transgenic maize inbred lines and hybrids with GFM Cry1A gene.","authors":"Y. Ying, Lin Xiao-hui, Kong Xiang-mei, Jia Zhi-lei, Lin Chun-Jing, L. Cong-cong, Liu Na, Sun Chuan-bo, Li De-pu","doi":"10.1017/S1479236209002654","DOIUrl":"https://doi.org/10.1017/S1479236209002654","url":null,"abstract":"Six transgenic maize lines with the fully modified gene GFM Cry1A were obtained and polymerase chain reaction (PCR) analysis of their T 5 generation plants indicated that foreign genes could be stably inherited. Three hybrids (Simi25 Bt , Tongdan24 Bt and Jidan209 Bt ) were created using the transgenic lines and combined with regular maize inbred lines. The results of resistance identification of the transgenic inbred lines and hybrids showed that the difference of resistance among transgenic lines was very significant; there also existed a difference among individuals of the same line and among the three hybrids. Compared with the control inbred line (CK), the average of four transgenic inbred lines in the number of tunnels per stalk, larval tunnel length per plant and length of surviving larvae were decreased by 70%, 80% and 70%, respectively. Enzyme-linked immunosorbent assay (ELISA) testing showed that the average of Bt Cry1A protein expression level for six transgenic inbred lines was 0.16% total protein and the highest expression level was 0.19%. At maturity, compared with the control variety (CK), the larval tunnel length per plant of the three hybrids (Simi25 Bt , Tongdan24 Bt and Jidan209 Bt ) was decreased by 39.97%, 36.20% and 53.83%, respectively, which was a decrease of 43.36% on average. The investigation of agronomic characters showed that there was no significant difference between the improved hybrids and the control in plant height, ear length, row number per ear, kernel number per row and 100-kernels weight. It is thought that the GFM Cry1A gene can be applied to improve resistance to corn-borer, and maize inbred lines with the Bt gene can be directly applied to conventional maize breeding.","PeriodicalId":236932,"journal":{"name":"Chinese Journal of Agricultural Biotechnology","volume":"23 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2009-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"121494313","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 : 2009-08-01DOI: 10.1017/S1479236209002642
Ma Jianjun, Yang Xiu-fen, Zeng Hongmei, Yuan Jing-jing, Qiu De-wen
N6, MS and BN mediums were adopted for tissue culture of Nipponbare (japanica) scutellum, and the effects of the three mediums on the cultrue were compared. Results indicated that, among the three kinds of culture mediums, NB medium worked best in the calli induction and gene transformation and was most suitable for tissue culture of Nipponbare scutellum. Based on this result, elicitor-encoding gene pemG1 from Magnaporthe grisea was introduced into the genome of Nipponbare via Agrobacterium-mediated method. And transgenic rice plants were obtained. The integration, transcription and expression of pemG1 in rice were confirmed by PCR, Northern blot and Western blot, respectively. Genetic analysis showed that the transgenes were segregated normally in the progenies.
{"title":"Medium selection in tissue culture of japonica rice cultivar Nipponbare and preparation of transgenic plants with an elicitor-coding gene from Magnaporthe grisea","authors":"Ma Jianjun, Yang Xiu-fen, Zeng Hongmei, Yuan Jing-jing, Qiu De-wen","doi":"10.1017/S1479236209002642","DOIUrl":"https://doi.org/10.1017/S1479236209002642","url":null,"abstract":"N6, MS and BN mediums were adopted for tissue culture of Nipponbare (japanica) scutellum, and the effects of the three mediums on the cultrue were compared. Results indicated that, among the three kinds of culture mediums, NB medium worked best in the calli induction and gene transformation and was most suitable for tissue culture of Nipponbare scutellum. Based on this result, elicitor-encoding gene pemG1 from Magnaporthe grisea was introduced into the genome of Nipponbare via Agrobacterium-mediated method. And transgenic rice plants were obtained. The integration, transcription and expression of pemG1 in rice were confirmed by PCR, Northern blot and Western blot, respectively. Genetic analysis showed that the transgenes were segregated normally in the progenies.","PeriodicalId":236932,"journal":{"name":"Chinese Journal of Agricultural Biotechnology","volume":"56 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2009-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"124865685","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 : 2009-08-01DOI: 10.1017/S1479236209990180
Fan Jia, Chen Ju-lian, Cheng Dengfa, Sun Jingrui
{"title":"Cloning and eukaryotic expression of olfaction-related Gqα-protein gene in English grain aphid ( Sitobion avenae )","authors":"Fan Jia, Chen Ju-lian, Cheng Dengfa, Sun Jingrui","doi":"10.1017/S1479236209990180","DOIUrl":"https://doi.org/10.1017/S1479236209990180","url":null,"abstract":"","PeriodicalId":236932,"journal":{"name":"Chinese Journal of Agricultural Biotechnology","volume":"104 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2009-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"115758525","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 : 2009-08-01DOI: 10.1017/S1479236209990234
GuoQing Xu, Dao-qing Gong, DongSheng Chu, D. Biao, Meng He
{"title":"Cloning and tissue expression of the adiponectin gene in geese.","authors":"GuoQing Xu, Dao-qing Gong, DongSheng Chu, D. Biao, Meng He","doi":"10.1017/S1479236209990234","DOIUrl":"https://doi.org/10.1017/S1479236209990234","url":null,"abstract":"","PeriodicalId":236932,"journal":{"name":"Chinese Journal of Agricultural Biotechnology","volume":"43 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2009-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"132003297","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 : 2009-08-01DOI: 10.1017/S1479236209002538
Zhang Linlin, W. Qi, Liao Rong-xi, Wang Yongjun, Mei Ru-hong
{"title":"Analysis, gene cloning and expression of two α-amylases from Bacillus cereus","authors":"Zhang Linlin, W. Qi, Liao Rong-xi, Wang Yongjun, Mei Ru-hong","doi":"10.1017/S1479236209002538","DOIUrl":"https://doi.org/10.1017/S1479236209002538","url":null,"abstract":"","PeriodicalId":236932,"journal":{"name":"Chinese Journal of Agricultural Biotechnology","volume":"21 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2009-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"121753198","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 : 2009-04-01DOI: 10.1017/S1479236209002514
C. Tao, Qian Guoliang, Yang Xiao-li, Ma Junyi, Hu Baishi, L. Fengquan
An efficient AHL ( N -acyl-homoserine lactone) bioassay strain, JZA1, of Agrobacterium tumefaciens was used to detect the AHL production from Acidovorax avenae subsp. citrulli [the pathogen causing bacterial fruit blotch (BFB) of melons], and the results showed that A. avenae subsp. citrulli produced a 3-O-C 8 -homoserine (HSL) type signal molecule. Gene aiiA , which could degrade AHL molecules, was transformed into A. avenae subsp. citrulli strain NJF10, creating strain NJF10- aiiA . The AHL production from NJF10- aiiA was significantly reduced compared with wild-type NJF10. Inoculation tests showed that NJF10- aiiA had an obvious reduction of virulence on watermelon fruits. Our finds showed that AHL production by A. avenae subsp. citrulli was related to its pathogenicity. This work might provide a novel way to control BFB by QS (quorum sensing) interference.
{"title":"Detection of a quorum sensing signal molecule of Acidovorax avenae subsp. citrulli and its regulation of pathogenicity","authors":"C. Tao, Qian Guoliang, Yang Xiao-li, Ma Junyi, Hu Baishi, L. Fengquan","doi":"10.1017/S1479236209002514","DOIUrl":"https://doi.org/10.1017/S1479236209002514","url":null,"abstract":"An efficient AHL ( N -acyl-homoserine lactone) bioassay strain, JZA1, of Agrobacterium tumefaciens was used to detect the AHL production from Acidovorax avenae subsp. citrulli [the pathogen causing bacterial fruit blotch (BFB) of melons], and the results showed that A. avenae subsp. citrulli produced a 3-O-C 8 -homoserine (HSL) type signal molecule. Gene aiiA , which could degrade AHL molecules, was transformed into A. avenae subsp. citrulli strain NJF10, creating strain NJF10- aiiA . The AHL production from NJF10- aiiA was significantly reduced compared with wild-type NJF10. Inoculation tests showed that NJF10- aiiA had an obvious reduction of virulence on watermelon fruits. Our finds showed that AHL production by A. avenae subsp. citrulli was related to its pathogenicity. This work might provide a novel way to control BFB by QS (quorum sensing) interference.","PeriodicalId":236932,"journal":{"name":"Chinese Journal of Agricultural Biotechnology","volume":"5 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2009-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"117102912","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 : 2009-04-01DOI: 10.1017/S147923620900254X
C. Jian-qiu, A. Yu-fang, Zhou Yifei, P. Da-ren, Xiong Guang-Ming, Guo Yu-chun, Pan Fei, Lin Bing-Hui
The testosterone-inducible regulator ( teiR ) gene was cloned from Comamonas testosteroni chromosomal DNA, and introduced into plasmids pKtac2 (containing a tac promoter) and pK18 to yield plasmids pKtac2- teiR and pK teiR100 . The recombinant plasmids were transformed into competent Escherichia coli HB101 and total protein was extracted to detect the TeiR protein expression level using enzyme-linked immunosorbent assay (ELISA). E. coli transformed by pKtac2- teiR and pK teiR100 produced 6.65 and 5.93 μg/mg of TeiR protein, respectively. Recombinant plasmids were also co-transformed into competent E. coli HB101 with plasmid p6 [containing hsdA gene (3α-HSD/CR, 3α-hydroxysteroid dehydrogenase/carbonyl reductase encoding gene)] to reveal the relationship between 3α-HSD/CR and TeiR by ELISA. The amounts of TeiR protein expressed by E. coli containing pKtac2- teiR and pK teiR100 were 5.94 μg/mg and 5.33 μg/mg, respectively, and these increased up to 6.81 μg/mg and 6.10 μg/mg after inducing with 1 mmol/l isopropyl-β- d -thiogalactoside (IPTG). Interestingly, 3α-HSD/CR protein expression level, after co-transformation with plasmids pKtac2- teiR and p6, was lower than that observed in the co-transformation with pK teiR100 and p6. The first co-transformation induced 1.20 μg/mg 3α-HSD/CR protein and the second 1.71 μg/mg. These values rose to 1.42 and 1.80 μg/mg, respectively, after treatment with 1 mmol/l IPTG. Our results proved that the tac promoter was more efficient than the lacZ promoter and that the teiR gene could act as an activator for hsdA gene expression.
{"title":"Characterization and heterologous expression of testosterone-inducible regulator from Comamonas testosteroni in Escherichia coli.","authors":"C. Jian-qiu, A. Yu-fang, Zhou Yifei, P. Da-ren, Xiong Guang-Ming, Guo Yu-chun, Pan Fei, Lin Bing-Hui","doi":"10.1017/S147923620900254X","DOIUrl":"https://doi.org/10.1017/S147923620900254X","url":null,"abstract":"The testosterone-inducible regulator ( teiR ) gene was cloned from Comamonas testosteroni chromosomal DNA, and introduced into plasmids pKtac2 (containing a tac promoter) and pK18 to yield plasmids pKtac2- teiR and pK teiR100 . The recombinant plasmids were transformed into competent Escherichia coli HB101 and total protein was extracted to detect the TeiR protein expression level using enzyme-linked immunosorbent assay (ELISA). E. coli transformed by pKtac2- teiR and pK teiR100 produced 6.65 and 5.93 μg/mg of TeiR protein, respectively. Recombinant plasmids were also co-transformed into competent E. coli HB101 with plasmid p6 [containing hsdA gene (3α-HSD/CR, 3α-hydroxysteroid dehydrogenase/carbonyl reductase encoding gene)] to reveal the relationship between 3α-HSD/CR and TeiR by ELISA. The amounts of TeiR protein expressed by E. coli containing pKtac2- teiR and pK teiR100 were 5.94 μg/mg and 5.33 μg/mg, respectively, and these increased up to 6.81 μg/mg and 6.10 μg/mg after inducing with 1 mmol/l isopropyl-β- d -thiogalactoside (IPTG). Interestingly, 3α-HSD/CR protein expression level, after co-transformation with plasmids pKtac2- teiR and p6, was lower than that observed in the co-transformation with pK teiR100 and p6. The first co-transformation induced 1.20 μg/mg 3α-HSD/CR protein and the second 1.71 μg/mg. These values rose to 1.42 and 1.80 μg/mg, respectively, after treatment with 1 mmol/l IPTG. Our results proved that the tac promoter was more efficient than the lacZ promoter and that the teiR gene could act as an activator for hsdA gene expression.","PeriodicalId":236932,"journal":{"name":"Chinese Journal of Agricultural Biotechnology","volume":"24 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2009-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"134397364","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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