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245 1 0 _aDevelopment of degenerate and specific PCR primers for the detection and isolation of known and putative chloroethene reductive dehalogenase genes
490 0 _vJournal of MicroBiological Methods, 56(1), p.107-118, 2004
520 3 _aDegenerate and specific PCR primers were designed for the detection of chloroethene reductive dehalogenases (CE-RDase), the key enzymes of chloroethene dehalorespiration, based on sequence information of three CE-RDases and three chlorophenol (CP)RDases. For the design of the degenerate primers, seven conserved amino-acid blocks identified with different bioinformatic tools were used. For one block degenerate, primers containing a 5V-consensus clamp region specific for CERDases and a 3V-end degenerate core region specific for RDases in general were designed using the Consensus-Degenerate Hybrid Oligonucleotide Primer (CDHOP)design method. Applying the degenerate primers to genomic DNA of Sulfurospirillum multivorans strain K, Dehalobacter restrictus strain PER-K23, and Desulfitobacterium sp. strain PCE1 led to the isolation of the known CE-RDase genes and three new genes encoding putative reductive dehalogenases that cluster with CE-RDases and not with CP-RDases. In addition, primers designed to be specific for the three known CE-RDase genes, namely pceA of S. multivorans, pceA of D. restrictus, and tceA of Dehalococcoides ethenogenes were successfully tested on genomic DNA of different chloroethene-dehalorespiring bacteria. Nested PCR using degenerate primers followed by a PCR with specific primers allowed a sensitive detection of only 102 copies per reaction.
650 1 4 _aREDUCTIVE DECHLORINATION
650 1 4 _aDEHALORESPIRATION
650 1 4 _aDEGENERATE PCR
650 1 4 _aBIOREMEDIATION
700 1 2 _aRegeard, Ch.
700 1 2 _aMaillard, J.
700 1 2 _aHolliger, C.
856 4 0 _uhttps://drive.google.com/file/d/1LIjEzneeQTUI8XDU331mgjZGGUBLtHce/view?usp=drivesdk
_zPara ver el documento ingresa a Google con tu cuenta: @cicy.edu.mx
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