从甘油富集菌宏基因组中克隆甘油脱水酶基因

Clonging the Genes Encoding Glycerol Dehydratase from Metagenomic DNA

采用一种简便快速的方法从经甘油富集培养的土样中提取出质量较好宏基因组DNA。然后以此DNA为模板,以扩增肺炎克雷伯氏菌、弗氏柠檬酸菌和丁酸梭菌甘油脱水酶基因的引物进行PCR,分别扩增出目的条带,并将其克隆至T载体中。进行测序分析显示,PCR扩增出来的片段与其引物相应的甘油脱水酶序列同源性分别达到99%,90%和99%。这表明克隆出来的这3个基因为相应的甘油脱水酶基因。

A pure metagenomic DNA was extracted from the soil by a simple, rapid method. From the metagenomic DNA, three different DNA fragments were successfully amplified using the primers of different glycerol dehydratase genes. And those fragments were inserted into T vector. After being sequenced, it was found that they were highly homologous to the reported glycerol dehydratase genes from Klebsiella pneumoniae, Citrobacter freundii and Clostridium butyricum. The sequences alignment shewed that these DNA fragments had 99 %, 90% and 99% identities to the strains above, respectively.