摘要
①目的 构建重组蜂毒肽的原核表达克隆。②方法 人工合成含特异性酶切位点的A ,B两条寡核苷酸片段 ,在Klenow酶作用下形成目的基因 ,用限制性内切酶HindⅢ ,XmnⅠ同时酶切目的基因和表达载体Pmal p2质粒 ,在T4连接酶的作用下构建两者的重组体 ,通过α 互补筛选出阳性克隆 ,并通过特异性酶切和测序分析进行鉴定。③结果 筛选出的阳性克隆为重组的蜂毒肽表达克隆。④结论 用分子生物学的方法重组构建的蜂毒肽基因表达克隆 ,是蜂毒肽蛋白表达及活性鉴定的基础。
Objective\ To construct mellitin gene expression clone. \ Methods\ Two fragments of manmade oligonucleotide,A and B, were made into target gene by Klenow enzyme . Then ,the target gene and prokaryotic expression vector Pmal p2 were restricted simultaneously and ligated by T4 DNA ligase. The recombinant plasmid was transformed into E.coli. DH5α. depending on the signs of α complementary. We selected the positive clone and it was confirmed by restriction enzyme and DNA sequencing. \ Results\ The positive clone was identified with recombinant mellitin gene clone. \ Conclusion\ The construction of recombinant mellitin gene expression clone is the basis of mellitin protein expression and activity identification . [
出处
《青岛大学医学院学报》
CAS
2001年第1期31-32,共2页
Acta Academiae Medicinae Qingdao Universitatis
基金
山东省教育厅科研基金资助项目!(J98K0 3 )
