CAJ | 학술논문

[目的]构建以pUC19质粒为基础可利用Xcm Ⅰ内切酶制备的T载体.[方法]化学合成2条含有双Xcm Ⅰ酶切位点的互补寡聚核苷酸链,经过变性、复性后克隆入pUC19质粒的HindⅢ和BamH1位点之间,通过Xcm Ⅰ酶切后得到一个线性化的带有3'末端突出一个T碱基的T载体.[结果]经TA克隆验证,制备的pUC19-HB-T载体对PCR产物的克隆率达到95%以上.[结论]所构建的pUC19-HB-T载体可用于PCR产物的克隆、测序及后续分子生物学操作.
[목적]구건이pUC19질립위기출가이용Xcm Ⅰ내절매제비적T재체.[방법]화학합성2조함유쌍Xcm Ⅰ매절위점적호보과취핵감산련,경과변성、복성후극륭입pUC19질립적HindⅢ화BamH1위점지간,통과Xcm Ⅰ매절후득도일개선성화적대유3'말단돌출일개T감기적T재체.[결과]경TA극륭험증,제비적pUC19-HB-T재체대PCR산물적극륭솔체도95%이상.[결론]소구건적pUC19-HB-T재체가용우PCR산물적극륭、측서급후속분자생물학조작.
[Objective] The paper aimed to construct a new T vector based on plasmid pUC19 digested with Xcm Ⅰ. [Method] Two complementary oligonucleotide chains containing two Xcm Ⅰ sites were synthesized. After denaturation and renaturation, the adaptor was cloned into plasmid pUC19 between the Hind Ⅲ and BamH I sites. The new plasmid, pUC19-HB-T vector, was digested with Xcm Ⅰ to derive a T-vector with 3' end overhanging a T base. [Result] The constructed pUC19-HB-T vector was efficient in cloning PCR products, with an efficiency of 95% at least,[Conclusion] A new Xcm Ⅰ-based pUC19-HB-T vector was constructed, which could be applied to cloning of PCR products and other microbiology operations.