CAJ | 학술논문

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Hind Ⅲ,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。
①목적구건중조봉독태적원핵표체극륭。②방법인공합성함특이성매절위점적A,B량조과핵감산편단，재Klenow매작용하형성목적기인，용한제성내절매Hind Ⅲ,XmnⅠ동시매절목적기인화표체재체Pmal-p2질립，재T4련접매적작용하구건량자적중조체，통과α-호보사선출양성극륭，병통과특이성매절화측서분석진행감정。③결과사선출적양성극륭위중조적봉독태표체극륭。④결론용분자생물학적방법중조구건적봉독태기인표체극륭，시봉독태단백표체급활성감정적기출。
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 .