Evidence for chimeric sequences formed during random arbitrarily primed PCR.
Chimeric sequences were observed to occur abundantly (48% of clones) during random arbitrarily primed polymerase chain reaction (RAP-PCR) experiments designed to examine differential expression of genes involved in metal resistance in sulfate-reducing bacteria (SRB). Some of the chimeric sequences were composed of sequence from a gene differentially expressed under the imposed conditions and a sequence of the 16S or 23S rRNA gene. The remainder were composed of two rRNA sequences. Experiments using PCR and genomic sequence analysis showed that the chimeric sequences were not due to a genetic mutation (e.g., recombination, transposition). As RAP-PCR has been widely used to identify differentially expressed genes, this observation may aid in our interpretation of RAP-PCR data.
Chang
In Seop
IS
Department of Botany and Microbiology, University of Oklahoma, Norman, OK 73019-0245, USA.
Ballard
Jimmy D
JD
Krumholz
Lee R
LR
D001483:Base Sequence; D017931:DNA Primers; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D016969:Desulfovibrio vulgaris / Q000737:chemistry / Q000235:genetics*; D005189:False Positive Reactions; D005798:Genes, Bacterial / Q000235:genetics; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics; D012335:RNA, Ribosomal / Q000737:chemistry / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics*; D017422:Sequence Analysis, DNA
D017931:DNA Primers; D004269:DNA, Bacterial; D012333:RNA, Messenger; D012335:RNA, Ribosomal; D011993:Recombinant Fusion Proteins