水牛Cdc42 cDNA的克隆与生物信息学分析

Cloning and Sequence Analysis of Cdc42 cDNA from Buffalo

通过RT-PCR扩增得到水牛细胞分裂周期蛋白42(Cdc42)cDNA序列,将扩增产物与pMD20-T载体连接,重组质粒经PCR、酶切鉴定后测序并进行生物信息学分析。结果表明:克隆得到的序列全长626bp,含有1个大小为576bp的开放阅读框,共编码191个氨基酸,相对分子质量为21.3×103,理论等电点为6.16;Cdc42ORF序列与Genbank上登载的黄牛、狗、小鼠和人的核苷酸序列同源性分别为99.65%、96.70%、94.97%和94.10%,具有很高的相似性;该蛋白为非跨膜蛋白,存在1个RHO结构域。

The objective of the study was to clone full-length cDNA of Cdc42（Cell division cycle 42） gene of buffalo and analyze this gene.RT-PCR were performed to clone Cdc42 cDNA from buffalo, and the cloned Cdc42 cDNA sequence had been sent to GeneBank and got a accession number：JN380079.The bioinformatics analysis results showed that the Cdc42 cDNA sequence was 626 bp length,containing an open reading frame of 576 bp,encoding 191 amino acids.Relative molecular weight was 21.3 kD,pI was 6.16.Further analysis results indicated that the homology of Cdc42 gene of buffalo are 99.65%, 96.70%, 94.97% and 94.10%, compared to cattle, dog, mouse and human, respectively; the protein was non-transmembrane protein, which include a RHO domain. These results set a strong basis for further research of the function of Cdc42 of buffalo.