【摘要】 根据已知的乙酰辅酶A羧化酶(ACCase)序列设计合成了1对引物,对ACCase基因的BC功能域进行扩增,所得产物与预期片段大小一致,约1.8kb。该片段与克隆载体PGEM TEase连接,转入感受态大肠杆菌DH5а中增殖。提取质粒进行PCR鉴定,将阳性克隆与原核表达载体PQE30分别用KpnⅠ和SalⅠ双酶切后回收目的片段进行连接,并转入感受态大肠杆菌M15中,所获重组质粒经过酶切、测序鉴定,证实含有目的片段,且连接、构建正确。更多还原

【Abstract】 In this study,primers P1 and P2 were designed according to the published DNA sequence of Acety-CoA Carboxylase and was used to amplify the BC structural gene of Acety-CoA Carboxylase by polymerase chain reaction(PCR).The PCR product was then cloned into the prokaryotic expression PGEM-T Ease vector,and the direct recombinants﹙PGEM-BC were obtained.In order to construct a prokaryotic expression vector containing BC structural gene of Acety-CoA Carboxylase double digestion KpnⅠand SalⅠ were performed to deal with BC and PQE30 respectively for the directional ligation of the two fragments.The following restriction endonuclease analysis has proved the correctness of the expression vector.更多还原