甲基营养菌MP688基因组完成图构建

Construct Genome Complete Map of Methylotrophic Bacteria MP688

目的:在Solexa测序建立的甲基营养菌MP688基因组精细图的基础上,构建MP688基因组完成图。方法:根据MP688基因组序列设计引物,进行常规PCR扩增,填补Scaffold内部缺口;利用BLASTN、BLAT软件分析预测6个Scaffold定位关系;通过组合PCR、长距离PCR等填充Scaffold间的物理缺口。结果:常规PCR填充Scaffold内部4个缺口;BLASTN、BLAT软件分析预测了Scaffold定位关系;组合PCR填充Scaffold间的2个物理缺口,长距离PCR填充其余4个物理缺口。结论:得到了MP688的基因组完成图,为MP688的吡咯喹啉醌生物合成途径的阐明和进一步的代谢工程育种奠定了基础。

Objective： In base of methylotrophic bacteria MP688 genome fineness map obtained by Solexa sequencing,we construct MP688 genome complete map.Methods： Using common PCR with primers based on MP688 genome sequence,we amplified the sequence gaps.Then we used BLASTN and BLAT software analysis to confirm the relationship between six Scaffolds.Finally,we used combination PCR and long distance PCR to fill the physical gaps.Results： We filled 4 sequence gaps by common PCR and confirmed the relationship between Scaffolds by BLASTN and BLAT software analysis,and filled 2 physical gaps by the combination PCR and the other 4 physical gaps by long distance PCR.Conclusion： We completed the MP688 genome map which lay a foundation to establish a strategy to elucidate the biosynthetic pathway and construct high pyrroloquinoline quinone producing strains by metabolic engineering.