人胎盘G-蛋白Rab3a cDNA的克隆

为了获取全长的小分子G-蛋白Rab3a，以用于研究Rab3a与其他蛋白相互作用关系，本实验以人胎盘总cDNA为模板，PCR扩增到人Rab3a cDNA全编码区。产物回收后克隆于质粒pYESTrp2的BamHI/XhoI位点，测序结果表明，本实验获得的Rab3a cDNA包含了起始和终止密码子。与PCR引物设计的参照Rab3a比较有5个核苷酸变异，与翻译的氨基酸序列完全一致。由此表明本实验获得的Rab3a cDNA可用于进一步研究。

Cloning of Rab3a cDNA from Human Fetal Placenta

In order to get small molecular G-protein Rab3a, which serves to further investigate the interaction between Rab3a and other proteins, we amplified the full coding region of Rab3a cDNA by polymerase chain reaction(PCR), using human placenta total cDNA as template. The PCR products were recovered from gel electrophoresis and cloned into BamHI/XhoI site of vector pYESTrp2. The result of sequencing indicated that Rab3a insertion fragment included its initiation and termination codons in 5′- and 3′-terminal, respectively. Compared with the referred Rab3a, the amplified Rab3a beard five nonsense nucleotide mutations, but the deduced amino acid sequence from the nucleotide sequence were completely the same as the referred Rab3a protein, which demonstrated that the cloned placenta Rab3a was suitable for further study.