Detection and quantification of phnE gene from oil-contaminated soil samples by competitive quantitative PCR.
The phnE gene encoding catechol 2,3-dioxygenase belonging to the meta-cleavage pathway was selected as the marker gene and was detected and quantified from soil samples by competitive quantitative PCR. A PCR primer pair was designed based on the phnE gene to amplify the target DNA bands and competitor DNA bands. The phnE gene was detected in two samples of three. In samples S1 and S2, the phnE gene copy number was 6.2 x 10(7)/g soil and 5.8 x 10(7)/g soil, respectively. But no phnE gene was detected in sample S3. The target DNA bands were extracted and expressed. The results confirmed that the target DNA bands were the native phnE genes.
State Key Laboratory of Microbial Technology, Shandong University, Jinan 250100, PR China.
Peng
P
Lui
Chang Kai
CK
Zhang
D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D050557:Catechol 2,3-Dioxygenase / Q000235:genetics*; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000032:analysis / Q000737:chemistry / Q000235:genetics*; D004787:Environmental Pollution; D008969:Molecular Sequence Data; D010578:Petroleum; D016133:Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA; D012987:Soil / Q000032:analysis; D012988:Soil Microbiology
D001426:Bacterial Proteins; D017931:DNA Primers; D004269:DNA, Bacterial; D010578:Petroleum; D012987:Soil; D050557:Catechol 2,3-Dioxygenase