| 双拷贝人α型心钠素基因的组建及大肠杆菌分泌型克隆与表达 |
| |
| 引用本文： | 周春燕,崔瑛琦,张丽霞,马康涛,王建平,曾桂超,张迺蘅,汤健,石松,王启松. 双拷贝人α型心钠素基因的组建及大肠杆菌分泌型克隆与表达[J]. 中国生物化学与分子生物学报, 1992, 8(1): 26-32 |
| |
| 作者姓名： | 周春燕 崔瑛琦 张丽霞 马康涛 王建平 曾桂超 张迺蘅 汤健 石松 王启松 |
| |
| 作者单位： | 北京医科大学生化教研室，北京医科大学生化教研室，北京医科大学生化教研室，北京医科大学生化教研室，北京医科大学心肺内分泌研究室，北京医科大学生化教研室，北京医科大学生化教研室，北京医科大学心肺内分泌研究室，复旦大学遗传研究所，复旦大学遗传研究所 北京 100083，北京 100083，北京 100083，北京 100083，北京 100083，北京 100083，上海，上海 |
| |
| 摘 要： | 将单拷贝人α心钠素基因3′端用Ban Ⅱ酶解除去包括终止密码在内的36个碱基对,代之以人工合成的含Glu-Lys-Phe-Glu连接片段与另一单拷贝人α心钠素基因的5′端串连成编码60肽的双拷贝心钠素基因,克隆于大肠杆菌分泌型表达载体pIN-Ⅲ-OmpA_2质粒中,表达生成60肽的双拷贝人α型心钠素衍生物,在信号肽的作用下分泌至胞膜间质并自动切割为60肽的外源基因产物。分子量约8K的表达产物用分子筛或超滤膜分离后再经HPLC纯化,表达产物具有明显的心钠素放免活性和舒张血管活性。
|
| 关 键 词： | 人心钠素 重组DNA 串联基因 分泌型表达质粒 |
| 收稿时间： | 1992-02-20 |
Recombination and Secretive Expression of Double-joined a-ANF Gene in E. coli. |
| |
| Zhou,Chun-yan Cui,Ying-qi Zhang,Li-xia Ma,Kang-tao Wang,Jian-ping Zeng,Gui-chao Zhang,Nai-heng Tang,Jian. Recombination and Secretive Expression of Double-joined a-ANF Gene in E. coli.[J]. Chinese Journal of Biochemistry and Molecular Biology, 1992, 8(1): 26-32 |
| |
| Authors: | Zhou Chun-yan Cui Ying-qi Zhang Li-xia Ma Kang-tao Wang Jian-ping Zeng Gui-chao Zhang Nai-heng Tang Jian |
| |
| Affiliation: | (Department of Biochemistry,Beijing Medical University,Beijing 100083) (Deparlment of Cardiopulmonary Endocrinology,BMU)Shi,Song Wang,Qi-song (Institute of Genetics,Fudan Univeristy |
| |
| Abstract: | A synthetic gene for a-hANF was reconstructed as a double joined gene,linked in tandem.The two copies in the gene were seperated by codons specifying a Glu-Lys-Phe-Glu linker with Glu residues flanking the authentic N and C termini of the 28-a.a.hormone.This linker is provided for enzymatic cleavage of expressed product by endoproteinaese Glu-c or Lys-c.The double joined gene fragment was cloned and expressed in) secretive expression vector PIN-*******III-OmpA2 in E.coli.The product was located in periplasmic space of the bacteria and purified by gel filtration and HPLC.The expressed protein after gently oxidized exhibited Vaso-dilatation activity similar to natural *********a-ANF. |
| |
| Keywords: | Human artrial natriuretic factor (ANF) Recombinant DNA Tandem gene Secretive expression vector |
| 本文献已被 CNKI 等数据库收录！ |
| 点击此处可从《中国生物化学与分子生物学报》浏览原始摘要信息 |
|
点击此处可从《中国生物化学与分子生物学报》下载全文 |
|
