Cloning of the gene for delta 5-3-ketosteroid isomerase from Pseudomonas testosteroni.
We have cloned an approx. 5-kb fragment of Pseudomonas testosteroni DNA containing the structural gene of delta 5-3-ketosteroid isomerase into the EcoRI site of the lambda gt11 genome. Escherichia coli infected with these recombinant phages produce a polypeptide which is recognized by antiserum raised against the purified isomerase. Four of the recombinant lambda gt11 clones contain significant levels of isomerase activity and produce an immunopositive polypeptide of the same apparent Mr as the native isomerase obtained from P. testosteroni. The approx. 5-kb fragment hybridizes to synthetic 21-mer and 17-mer oligodeoxynucleotide mixtures corresponding to the 5' and 3' regions, respectively, of the expected nucleotide sequence of the gene.
Department of Biological Chemistry, School of Medicine, University of California, Davis 95616.
K Y
KY
Choi
W F
WF
Benisek
D001483:Base Sequence; D003001:Cloning, Molecular; D005796:Genes; D005798:Genes, Bacterial; D007535:Isomerases / Q000235:genetics*; D009693:Nucleic Acid Hybridization; D011549:Pseudomonas / Q000201:enzymology / Q000235:genetics*; D013251:Steroid Isomerases / Q000235:genetics*
D007535:Isomerases; D013251:Steroid Isomerases; C022660:steroid delta-isomerase