Open AccessThe triazine hydrolase gene trz N from Nocardioides sp. strain C190: Cloning and construction of gene‐specific primers
Author(s) -
Mulbry Walter W.,
Zhu Hong,
Nour Sarah M.,
Topp Edward
Publication year - 2002
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2002.tb10989.x
Subject(s) - transposase , biology , microbiology and biotechnology , gene , nucleic acid sequence , rhodococcus , escherichia coli , actinomycetales , molecular cloning , transposable element , peptide sequence , biochemistry , streptomyces , genetics , bacteria , mutant
Using oligonucleotides derived from the N‐terminal sequence of a triazine hydrolase from Nocardioides sp. strain C190, two DNA fragments containing trz N were cloned into Escherichia coli and their nucleotide sequences were determined. The 456‐amino acid polypeptide predicted from the 1356‐bp trz N ORF displayed significant similarity to triazine hydrolases from Pseudomonas and Rhodococcus isolates and belonged to the same amidohydrolase family. The trz N gene was flanked by two DNA sequences possessing 57 and 69% identity, respectively, at the protein level to Rhodococcus erythropolis sequences for a transposase and a transposase helper protein. Amplification primers specific to trz N were tested in soils inoculated with strain C190. The results demonstrated that the primers were specific to trz N, and could detect populations at 10 8 cfu g −1 soil using 250‐mg soil samples.