湖北白猪胃蛋白酶原A基因cDNA克隆与序列分析

根据Genebank公布的J04601猪胃蛋白酶原A序列,进行了引物的设计和筛选,通过RT-PCR获得了湖北白猪的胃蛋白酶原A的cDNA序列全长,共1 227 bp(已提交Genebank,登录号EF108301),其具有一个长度为1 158 bp的读码框,编码385个氨基酸,其中包含15个氨基酸组成的信号肽,成熟的胃蛋白酶原A由370个氨基酸残基组成.与J04601比较,核苷酸和氨基酸相似性分别为99.3%和98.7%.经PredictProtein分析表明,核苷酸和氨基酸的改变没有造成蛋白质二级结构和功能的变化;并对胃蛋白酶原A基因密码子使用频率和mRNA的二级结构进行了分析,为其载体和宿主的选择以及基因的改造奠定了分子生物学基础.

Cloning and Sequence Analysis of cDNA for Pepsinogen A from Hubei White Swine

According to the sequence J04601 in Genebank,the primers were designed and flitrated.The Hubei White swine pepsinogen A cDNA was cloned by RT-PCR,which were 1 227bp in length(submitted in Genebank,the accession number is EF108301.),including a ORF(1 158bp),encoding 385 amino acids.The NH2-terminal region present a signal peptide consisted of fifteen amino acids,there are 370 amino acids in the mature protein.In the mature-protein regions,EF108301 sshowed 99.3% and 98.7% identical nucleotide and amino acid residues with J04601.PredictProtein analysis shows that the alteration of nucleotide and amino acid did not change the secondary structure and of the protein.The analysis of codon usage and mRNA secondary structure was benefited for the further research on the choice of vector,host and,the reconstruct of gene.