猪CISD1基因电子克隆与验证

对通过抑制差减杂交技术所筛选的猪前脂肪细胞诱导分化后差异表达的CISD1基因的EST序列为探针进行电子克隆,用克隆序列设计引物,通过RT-PCR、克隆测序、同源比对证实所获序列为猪CISD1基因的mRNA序列,并将序列提交到Genebank,登录号为GQ913654。所获猪CISD1基因的mRNA序列全长为631 bp,CDS包含321 bp,由其编码的CISD1蛋白包含106个氨基酸残基,理论等电点为9.20,属脂溶性不稳定蛋白。猪CISD1蛋白的1~23位氨基酸可能是信号肽,53~91位氨基酸形成CDGSH锌手指域,为该蛋白的重要功能域,猪CISD1蛋白的三级结构与人CISD1蛋白相似度为89.333%。

Electronic Cloning and Verification of Porcine CISD1 Gene

The Expressed Sequence Tag(EST) sequence of porcine CISD1 gene was obtained from the Suppressive Subtractive Hybridization(SSH) libraries,which construction of the induced differentiation of porcine preadipocytes.This EST sequence was used as a probe to predict the full-length cDNA sequence of porcine CISD1 gene using an in silico approach.The cDNA sequence of porcine CISD1 gene was obtained using a RT-PCR method,which has been deposited to NCBI-GenBank with the accession number DQ913654.The cDNA sequence of porcine CISD1 gene was 631 bp long,which containing a 321 bp-coding sequence(CDS) encode to 106 amino acids.The physical and chemical properties of porcine CISD1 protein were also predicted using an in silico approach.The results indicated that the CISD1 protein belongs to the unstable and liposoluble protein family(PI=9.2).The CISD1 protein also contains a signal peptide from 1st to 23th amino acid and a ZnF-CDGSH domain from 53th to 91th.The tertiary structure of porcine CISD1 has the high similarity(89.33%) to human CISD1 protein.