[Cloning of genes coding for 3-alpha-hydroxysteroid dehydrogenase and for (3-17)-beta-hydroxysteroid dehydrogenase from Pseudomonas testosteroni].
A genomic library of Pseudomonas testosteroi total DNA constructed from SauIIIA digests ligated to a lambda gt11 vector was probed with different polyclonal antibodies raised against purified 3 alpha-HSD and (3 beta-17 beta)-HSD. Two different clones reacting with one antibody were selected. The clone reacting with (3-17)beta-HSD antibody contained a 2,661-base pair insert and was found to contained an open reading frame of 765 base pair that corresponds to a protein of 254 amino-acid residues. A 1,492-base pair was inserte in pBR 322 plasmid vector; the recombinant bacterie over expressed the (3-17)beta-HSD gene. The clone reacting with 3 alpha-HSD antibody contained a 1746 base pair insert which contained an open reading frame of 696 base pairs that corresponds to a protein of 231 amino-acid residues. A search for homologous proteins was performed. Distant similarities were found between (3-17)beta-HSD and members of the short-chain alcool dehydrogenase (SCAD) family but no similarity was observed between 3 alpha-HSD and proteins of this family.
Floch
H H
HH
Département de Biochimie et de Biologie moléculaire, Faculté de Médecine, Brest.
Abalain
J H
JH
Di Stefano
S
S
Carré
J L
JL
Abalain-Colloc
M L
ML
D015096:3-Hydroxysteroid Dehydrogenases / Q000235:genetics*; D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004251:DNA Transposable Elements; D006913:Hydroxysteroid Dehydrogenases / Q000235:genetics*; D066298:In Vitro Techniques; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D011549:Pseudomonas / Q000201:enzymology*; D017386:Sequence Homology, Amino Acid
D004251:DNA Transposable Elements; D015096:3-Hydroxysteroid Dehydrogenases; D006913:Hydroxysteroid Dehydrogenases
Clonage des gènes codant pour la 3 alpha-hydroxystéroïde déshydrogénase et pour la (3-17)beta-hydroxystéroïde déshydrogénase de Pseudomonas testosteroni.