豇豆胰蛋白酶抑制剂基因的克隆

Cloning of cowpea trypsin proteinase inhabitor gene

利用 RT- PCR的方法 ,从豇豆种子、子叶及成熟叶片中分别提取 RNA,反转录后以该基因两端的保守序列为引物进行 PCR扩增 ,PCR产物与 p UCm- T载体连接进行克隆 ,蓝白筛选重组子 ,经测序 ,其核苷酸同源性高达 1 0 0 % ,蛋白质同源性达 91 %。本试验同时对该基因的表达时期差异进行了大致检测 ,结果是种子的表达量最高 ,子叶其次 。

Total RNA was isolated from seeds,cowpea cotyledons and matured leaves.The CpTI gene was amplified by PCR after reverse transcription with the conserved sequence acted as primers.PCR products was cloned with pUCm T vector.After sequence,the nucleotides acids homology was 100%,the proteines homology was 91%.The gene expression period difference was appromiatly detected with the highest expression volume at the seeds periods,the cotyledons was second,and the matured leaves was the lowest.