The triazine hydrolase gene trzN from Nocardioides sp. strain C190: cloning and construction of gene-specific primers.
Using oligonucleotides derived from the N-terminal sequence of a triazine hydrolase from Nocardioides sp. strain C190, two DNA fragments containing trzN were cloned into Escherichia coli and their nucleotide sequences were determined. The 456-amino acid polypeptide predicted from the 1356-bp trzN ORF displayed significant similarity to triazine hydrolases from Pseudomonas and Rhodococcus isolates and belonged to the same amidohydrolase family. The trzN 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 trzN were tested in soils inoculated with strain C190. The results demonstrated that the primers were specific to trzN, and could detect populations at 10(8) cfu g(-1) soil using 250-mg soil samples.
Mulbry
Walter W
WW
U.S Department of Agriculture, Agricultural Research Service, Beltsville, MD 20705-2350, USA. mulbryw@ba.ars.usda.gov
Zhu
Hong
H
Nour
Sarah M
SM
Topp
Edward
E
D000192:Actinomycetales / Q000201:enzymology* / Q000235:genetics*; D000595:Amino Acid Sequence; D001280:Atrazine / Q000378:metabolism; D001426:Bacterial Proteins; D001673:Biodegradation, Environmental; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics*; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D006540:Herbicides / Q000378:metabolism; D006867:Hydrolases / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA; D019895:Transposases / Q000235:genetics / Q000378:metabolism
D001426:Bacterial Proteins; D017931:DNA Primers; D006540:Herbicides; D019895:Transposases; D006867:Hydrolases; C448782:trzN protein, Nocardioides sp. C190; D001280:Atrazine