Analysis of mer Gene Subclasses within Bacterial Communities in Soils and Sediments Resolved by Fluorescent-PCR-Restriction Fragment Length Polymorphism Profiling.

Applied and Environmental Microbiology
K D Bruce

Abstract

Bacterial mer (mercury resistance) gene subclasses in mercury-polluted and pristine natural environments have been profiled by Fluorescent-PCR-restriction fragment length polymorphism (FluRFLP). For FluRFLP, PCR products were amplified from individual mer operons in mercury-resistant bacteria and from DNA isolated directly from bacteria in soil and sediment samples. The primers used to amplify DNA were designed from consensus sequences of the major subclasses of archetypal gram-negative mer operons within Tn501, Tn21, pDU1358, and pKLH2. Two independent PCRs were used to amplify two regions of different lengths (merRT(Delta)P [ca. 1 kb] and merR [ca. 0.4 kb]) starting at the same position in merR. The oligonucleotide primer common to both reactions (FluRX) was labelled at the 5(prm1) end with green (TET) fluorescent dye. Analysis of the mer sequences within databases indicated that the major subclasses could be differentiated on the basis of the length from FluRX to the first FokI restriction endonuclease site. The amplified PCR products were digested with FokI restriction endonuclease, with the restriction digest fragments resolved on an automated DNA sequencing machine which detected only those bands labelled with the fluores...Continue Reading

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Citations

Feb 7, 2003·Applied and Environmental Microbiology·Christopher B BlackwoodEldor A Paul
Aug 30, 2001·Applied and Environmental Microbiology·W H EschenfeldtM I Donnelly
Jul 11, 2013·World Journal of Microbiology & Biotechnology·N WatanarojanapornN Teaumroong
Jun 4, 2014·Indian Journal of Microbiology·Om PrakashYogesh S Shouche
Jan 27, 2021·Environmental Science and Pollution Research International·Jofre HerreroJosé M Carmona
Aug 3, 2001·International Journal of Food Microbiology·G Giraffa, E Neviani
Aug 7, 2021·Environmental Science and Pollution Research International·Jofre HerreroJosé M Carmona

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