Exploration of GC-rich Gene PCR System from Burkholderia pyrrocinia JK-SH007

GC-rich gene PCR system from biocontrol strain Burkholderia pyrrocinia JK-SH007 was established. The genome of strain JK-SH007 was processed for 5 min at 100 ℃. The total volumes of PCR were 10 μL. The PCR reactions added high-fidelity LA Taq (Mix) polymerase and different DMSO, 2×GC bufferⅠ, 2×GC bufferⅡ, respectively. Then the producing chitinase gene ( bcc1 , GC content: 71.1%) was amplified by the appropriate PCR procedures. And on this basis, the best PCR system was adopted for producing ACC deaminase gene (acdS, GC content: 66.86%), endoglucanase gene (GC content: 74.71%), tryptophan monooxygenase gene (iaaM, GC content: 68.62%) and siderophore biosynthesis-related gene (cepR, GC content: 64.44%), respectively. The amplified product was cloned into the PMDⓇ19-T vector and then transformed into E.coli JM-109 for sequencing. The result showed that the PCR system added 10% DMSO could be successfully amplify bcc1 gene (2 484 bp) and be equally applicable to acdS gene, endoglucanase gene, iaaM gene and cepR gene. Moreover, the sequencing results were in complete accord with the expected sequences. So this PCR system is a wide-range, simple, high quality, low-cost and effective way for amplification the GC-rich genes. All of these provide an important reference for the cloning of GC-rich genes of Burkholderia cepacia complex strains and others.