利用一碳化合物(C1)细菌的分离及基因组文库的构建

Isolation of the C1-utilization Ability Bacteria and Construction of Their Genomic Library

从污水样品中筛选到能利用甲醇的菌株B,经16SrDNA测序分析鉴定为肺炎克氏杆菌(Klebsiella pneumo-niae)。甲醛耐受能力的测试表明该菌对甲醛具有较强耐受能力,能在含有8￣15mmol/L甲醛的LB培养基上生长。Southern杂交分析说明这菌株的基因组中有甲基营养菌6-磷酸己酮糖合成酶(HPS)和6-磷酸果糖异构酶(PHI)基因的同源序列。本研究以pUC118为载体构建了基因组文库,进一步检测结果说明所构建的基因组文库质量符合要求。

The strain B with CH3OH-utilization ability was isolated from sewage samples. By sequence analyses for 16S rDNA, the isolated strain was identified as Klebsiella pneumoniae. The test with HCHO showed it was well tolerant to HCHO and could grow on LB medium with 8-15 mmol/L HCHO. Southern hybridization analyses revealed that the orthologs of 3- Hexulose-6-phosphate synthase （HPS） and 6-phospho-3-hexuloisomerase （PHI） genes were presented in the genomes of the isolated strain. Using pUC118 as the vector, genomic DNA library was constructed for the isolated strain. The further analyses showed that the quality of the library was met the requirement for gene screening.