[Construction of the pUCD-recA recombinate luminescence bacterium to evaluate the genetoxicty of environment pollutants].
Recombinate luminescence bacteria have the important role in evaluating water toxicity. A recombinate luminescence bacteria vector pUCD-recA was constructed to investigate the general toxicity of pollutants. The gene of recA amplified by PCR from W3110 was cloned into pGEM-T easy vector and sequencing. The correct PCR product and pUCD615 vector were digested with BamH I, EcoR I, then be fused and imported into JM109 with electrotransformation. Several clones were selected and identificated by PCR and sequencing. The result revealed that the length of the recA fragment was 293 bp. When it was sequenced and blasted with the recA in GenBank, the homology of the sequences reached 99% indicating the amplified result correct. The sequencing result of the fragment fused with pUCD615 revealed that the gene of recA had been inserted into the multiple clone site correctly, and the insert direction and reading frame were also exactly. Transforming the pUCD-recA vector to E. coli RFM443, then added the genetoxic pollutants and observed the responding effect. The effect of MMC inducing the pUCD-recA strain was best with the dose only of 0.01 mg/L, while need the dosage of MNNG reached 50-100 mg/L.
Huang
Xin-Xin
XX
Division of Water Environment, Department of Environmental Science and Engineering, Tsinghua University, Beijing 100084, China. xxhuang@tsinghua.edu.cn
He
Miao
M
Luo
Hong
H
Shi
Han-Chang
HC
Cai
Qiang
Q
D001426:Bacterial Proteins / Q000235:genetics; D001483:Base Sequence; D004785:Environmental Pollutants / Q000032:analysis*; D004926:Escherichia coli / Q000235:genetics*; D049449:Luminescence; D016685:Mitomycin / Q000032:analysis; D008969:Molecular Sequence Data; D009152:Mutagenicity Tests / Q000379:methods; D011938:Rec A Recombinases / Q000235:genetics* / Q000378:metabolism; D011995:Recombination, Genetic / Q000235:genetics*
D001426:Bacterial Proteins; D004785:Environmental Pollutants; D016685:Mitomycin; D011938:Rec A Recombinases