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| 睾丸酮丛毛单胞菌活化因子基因的质粒载体构建及表达 | |
| 引用本文： | 戴艺民,潘大仁,Guangming Xiong,吴明霞,周以飞.睾丸酮丛毛单胞菌活化因子基因的质粒载体构建及表达[J].农业生物技术学报,2006,14(3):416-422. |
| 作者姓名： | 戴艺民 潘大仁 Guangming Xiong 吴明霞 周以飞 |
| 作者单位： | 1. 福建农林大学作物学院,福州,350002;福建省农业科学院甘蔗研究所,漳州,363005 2. 福建农林大学作物学院,福州,350002 3. Institute of Toxicology and Pharmacology for Natural Scientces, University Medical School, Schleswig-Holstein, Campus Kiel, Brunswiker Str. 10, 24105 Kiel, Germany |
| 基金项目： | 福建省科技厅科研项目;福建省发展与改革委员会资助项目 |
| 摘 要： | 提取睾丸酮丛毛单胞菌(Comamonas testosteroni)的基因组DNA，利用PCR扩增activator基因，将扩增产物克隆到pKtac1(含强启动子tac)中，构建重组质粒pKtac1-act1和 pKtac1-act2。经酶切分析和测序，鉴定出正确的重组质粒pKtac1-act1和 pKtac1-act2。将重组质粒转化入大肠杆菌(Escherichia coli)HB101中，用酶联免疫分析方法测定细菌总蛋白质中的activator表达量；将pAX1(含3α-HSD/CR基因)分别和重组质粒(pKtac1-act1和pKtac1-act2)一起转化入E. coli HB101中，测定细菌总蛋白质中的3α-HSD/CR和activator的表达量。结果表明：重组质粒能明显提高activator的表达；pAX1与重组质粒共转化的蛋白质粗提物中，activator和3α-HSD/CR的表达量都显著提高。 |
| 关 键 词： | 睾丸酮丛毛单胞菌 活化因子 3α-羟基类固醇脱氢酶/碳酰基还原酶 基因表达 |
| 文章编号： | 1006-1304（2006）03-0416-06 |
| 收稿时间： | 2005-2-21 |
| 修稿时间： | 2006-3-30 |
Construction and Over-expression of the Activator Gene from Comamonas testosteroni |
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| DAI Yi-min,PAN Da-ren,Guangming Xiong,WU Ming-xia,ZHOU Yi-fei.Construction and Over-expression of the Activator Gene from Comamonas testosteroni[J].Journal of Agricultural Biotechnology,2006,14(3):416-422. | |
| Authors: | DAI Yi-min PAN Da-ren Guangming Xiong WU Ming-xia ZHOU Yi-fei |
| Institution: | 1. College of Crop Science, Fujian Agriculture and Forestry University, Fuzhou 350002, China; 2.Sugarcane Research Institute, Fujian Academy of Agricultural Sciences, Zhangzhou 363005, China; 3. Institute of Toxicology and Pharmacology for Natural Scientists, University Medical School Schleswig-Holstein, Campus Kiel, Brunswiker Str. 10, 24105 Kiel Germany |
| Abstract: | The activator gene from Comamonas testosteroni was amplified by PCR from the genomic DNA of C. testosteroni. The PCR products were cloned into plasmid pKtac1 (containing tac promoter), and the recombinant plasmids pKtac1-act1 and pKtac1-act2. were obtained and identified by the analysis of restriction enzymes and DNA sequencing and then respectively transformed into Escherichia coli HB101. The total cell lysate of the host bacteria was extracted to detect the quantity of activator using ELISA. Moreover, the recombinant plasmids were also co-transformed with plasmid pAX1 (containing 3α-HSD/CR gene) into E.coli HB101 respectively. The total cell lysate of the host bacteria was also extracted to detect the quantity of activator and 3α-HSD/CR using ELISA. Moreover, the results indicated that the recombinant plasmids can improve the expression of activator; and the expressions of activator and 3α-HSD/CR can be also improved by plasmid pAX1 co-transformed with recombinant plasmids. |
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