甲基营养菌MP681基因组测序文库的构建

目的:建立甲基营养菌MP681基因组文库,用于鸟枪法测序。方法:提取MP681基因组DNA,经超声随机片段化及T4 DNA聚合酶末端修平处理后,与经SmaⅠ酶切、小牛肠碱性磷酸酶(CIP)去磷酸化处理的pUC19载体连接,电击转化大肠杆菌DH5α感受态,并通过末端双向测序对文库质量进行评价。结果:分别构建了2～4 kb和4～6 kb基因组文库,电泳结果显示插入片段长度与预期符合,文库库容均在10万以上。结论:构建了插入片段大小和库容符合要求的甲基营养菌MP681全基因组鸟枪法2～4 kb、4～6 kb测序文库。

Genome Sequencing Libraries Construction of a Methylotrophic Bacteria Strain MP681

Objective： To construct methylotrophic bacteria strain MP681 genomic library for shotgun sequencing.Methods： The MP681 genome DNA was extracted and degradated with soniciation to random fragment.It was treated with T4 DNA polymerase and ligated to pUC19 vector which was digested by SmaⅠ and CIP.The ligation product was transformed into E.coli DH5α competent cell by electroporation.The genome libraries were accessed by double-ended sequencing.Results： The 2～4 kb and 4～6 kb genome libraries were constructed and the nucleic acid electrophoresis result indictaed that the length of insert fragment was enough,moreover the library capacity are both greater than 100 000.Conclusion： The 2～4 kb and 4～6 kb genome libraries were successfully constructed which the length of insert fragment and the capacity are both qualified.