遗传 ›› 1996, Vol. 18 ›› Issue (5): 27-30.

• 论文 • 上一篇    下一篇

中国仓鼠卵细胞二氢叶酸还原酶基因在大肠杆菌中的高效表达

蔡勤1; 徐晋麟2 CAI Qin1;XU Jin-Lin2   

  1. 1.中国科学院上海生物工程研究中心, 上海 200233 2.上海交通大学生物科学与技术系,上海 200240 1.Shanghai Research Center of Biotechnology,Academica Sinica,Shanghai 200233 2.Department of Biology Science and Technology of Shanghai JiaoTong University,Shanghai 200240
  • 收稿日期:1900-01-01 出版日期:1996-10-10 发布日期:1996-10-10

High-level Expression of CHO Dihydrofolate Reductase Gene in Escherichia coli

  • Received:1900-01-01 Online:1996-10-10 Published:1996-10-10

摘要: 本工作采用PCR法将哺乳动物表达载体pSVA75上的中国仓鼠卵细胞(CHO)二氢叶酸还原酶基因扩增后插入到原核生物高效表达质料pBV221中,构建成胞内表达载体pDHFR-1,经转化大肠杆菌(E.coli DH5α)后获高效表达,表达量占总蛋白量的12.2%。利用这种方法不仅为深入研究该酶的结构和功能的关系及其新生肽链的折叠提供丰富的实验材料,而且也有希望成为较好的外源DNA原核扩增表达系统。
Abstract:The technique of PCR was used to amplify the Chinese Hamster Ovary DHFR gene in the mammal expression vector pSVA75,and the amplified fragments were inserted into the vector pBV221,to construct the vector pDHFR-1.The CHO dhfr was fransformaed into E.coli DH5α and high-level expressed.The scanning results(scanned by LKB Enhanced Laser Densitometer)showed that the expression level had reached 12.2% of the total cellular proteins.Therefore,this provides an abundant experimental material for studying the relationship between the protein construct and function,and for studying the biology of the nascent peptide chain during biosynthesis.This pathway may be a good expression system of prokaryocyte for amphifing eukaryotic gene.

关键词: 二氢叶酸还原酶, CHO, PCR, pBV221, 高效表达
Key words,
RFLP, 紫色种皮, 基因定位, 水稻

Key words: Purple pericarp Gene mapping RFLP Rice