CLONING, DNA SEQUENCING AND EXPRESSION OF (3-17)BETA HYDROXYSTEROID DEHYDROGENASE FROM PSEUDOMONAS-TESTOSTERONI

被引：39

作者：

ABALAIN, JH ^{[1
]}

DISTEFANO, S ^{[1
]}

AMET, Y ^{[1
]}

QUEMENER, E ^{[1
]}

ABALAINCOLLOC, ML ^{[1
]}

FLOCH, HH ^{[1
]}

机构：

[1] FAC MED BREST,BACTERIOL LAB,F-29279 BREST,FRANCE

来源：

JOURNAL OF STEROID BIOCHEMISTRY AND MOLECULAR BIOLOGY | 1993年 / 44卷 / 02期

关键词：

D O I：

10.1016/0960-0760(93)90020-W

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

We describe the cloning, sequencing and overexpression of the (3-17)beta hydroxysteroid dehydrogenase gene of Pseudomonas testosteroni. A genomic library of Ps. testosteroni total DNA constructed from SauIIIA digests ligated to a lambdagt11 vector was probed with polyclonal antibody raised against purified enzyme. Subclones derived from a recombinant phage containing a 2661-base-pair insert were sequenced and found to contain an open reading frame of 765 base pairs that corresponds to a protein of 254 amino acid residues. A 1492-base-pair fragment was inserted into pBR322 plasmid vector and used to construct a strain of E. coli HB101 that overexpressed the steroid dehydrogenase gene.

引用

页码：133 / 139

页数：7

共 27 条

[1] HUMAN PLACENTAL 17-BETA-HYDROXYSTEROID DEHYDROGENASE IS HOMOLOGOUS TO NODG PROTEIN OF RHIZOBIUM-MELILOTI [J].