The triazine hydrolase gene trzN from Nocardioides sp. strain C190: Cloning and construction of gene-specific primers

• Published in: FEMS microbiology letters 2002-01, Vol.206 (1), p.75-79
• Main Authors: Mulbry, Walter W., Zhu, Hong, Nour, Sarah M., Topp, Edward
• Format: Article
• Language: English
• Subjects: Actinomycetales - enzymology; Actinomycetales - genetics; Agrochemicals products; Amino Acid Sequence; Atrazine - metabolism; Atrazine biodegradation; atrazine chlorohydrolase; Bacterial Proteins; Biodegradation, Environmental; Biodeterioration. Biofouling; Biological and medical sciences; Biotechnology; Cloning, Molecular; DNA; DNA fragmentation; DNA Primers - genetics; Escherichia coli; Escherichia coli - enzymology; Escherichia coli - genetics; Fundamental and applied biological sciences. Psychology; genes; Herbicides - metabolism; Hydrolases - genetics; Hydrolases - metabolism; Industrial applications and implications. Economical aspects; Molecular Sequence Data; Nocardioides; oligonucleotides; open reading frames; Other agrochemicals used in plant protection (herbicides, fungicides, etc.); Phytopathology. Animal pests. Plant and forest protection; Pollution effects and side effects of agrochemicals on crop plants and forest trees. Other anthropogenic factors; Pollution effects. Side effects of agrochemicals; polypeptides; Pseudomonas; Rhodococcus erythropolis; s-Triazine hydrolase; Sequence Analysis, DNA; soil; soil sampling; Transposase; Transposases - genetics; Transposases - metabolism; triazine hydrolase; triazines; trzN gene
• ISSN: 0378-1097; 1574-6968
• DOI: 10.1016/S0378-1097(01)00517-1

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.