海南原鸡CDC42 cDNA的克隆及其生物信息学分析

Cloning and Bioinformatics Analysis of CDC42 cDNA from Hainan Junglefowl

以海南原鸡外周血白细胞总RNA为模板,利用RT-PCR技术,扩增海南原鸡细胞分裂周期蛋白42(Cell division cycle42,CDC42)编码区,将扩增产物与pMDTM20T载体连接,重组质粒鉴定正确后,进行序列特征、同源性、编码蛋白质理化性质及结构预测等生物信息学分析。结果表明,海南原鸡CDC42的CDS序列长度为576bp,编码191个氨基酸;与鸡、人、牛、褐家鼠、斑马鱼的核苷酸序列同源性分别为100%、88.54%、88.19%、86.98%、82.12%;该基因编码的蛋白质相对分子质量约为21259,理论等电点为6.16;编码蛋白为非跨膜蛋白,在其二级结构中,α-螺旋占27.23%,β-折叠占25.65%,无规卷曲占47.12%。

Cell division cycle 42 （CDC42）cDNA of Hainan Junglefowl was amplified by RT-PCR and the sequence was analyzed. The bioinformatics analysis results indicated that CDC42 cDNA had an open reading frame contained 576 bp and encodied 191 amino acids; compared to Callus gallus, Homo sapiens, Bos taurus, Rattus norvegicus and Danio rerio, nucleotide sequence homology of CDC42 was 100%, 88.54%, 88.19%, 86.98%, 82.12%,respectively;relative molecular weight was 21 259, pI was 6.16; the encoded protein was non-transmembrane protein, which contained 27.23% α-helix,25.65% β-meander,47.12% random coil.