人Midkine基因启动子片段的克隆

目的克隆人Midkine（MK）启动子基因2 335bp片段.方法自健康人血中提取人类基因组DNA作为模板,聚合酶链式反应（PCR）技术克隆人MK启动子基因2 335bp片段.琼脂糖凝胶电泳初步鉴定PCR产物,切取目的条带纯化并将其克隆入p GEM T载体中.选取阳性克隆进行DNA测序.结果电泳结果显示PCR产物包含有大小约为2 300 bp和1 400 bp的条带各1条;较大片段的测序结果显示,其与Genbank中的人MK启动子片段吻合.结论本研究成功克隆了人MK启动子基因2 335 bp片段.

Cloning of human midkine promoter gene fragment

[Objective] To clone the 2 335 bp fragment(-2 285/50) of human midkine promoter gene.[Methods] As a template,the human genome DNA was extracted from healthy human blood firstly.And then,the polymerase chain reaction(PCR) was applied to clone the 2 335 bp fragment of human midkine promoter gene.After primary identification with agarose gel electrophoresis,the corresponding DNA fragment was purified and cloned into p GEM T vector.Finally,the recombinant plasmid was comfirmed with DNA sequencing technique.[Results] Besides of a fragment about 1 400 bp,another fragment about 2 300 bp was obtained by PCR.This DNA fragment was in accordance with the human midkine promoter dominantly with 8 mutant base pairs.[Conclusions] We have got the 2 335 bp fragment of human midkine promoter successfully.