猪CTSZ基因的克隆及真核表达载体的构建

摘要

Cathepsin Z gene (CTSZ) affect many traits of pigs, such as the growth and meat quality.It is an important candidate gene.The whole CDS sequence of CTSZ gene was cloing by RT-PCR applied to the longissimus dorsi of landrace.Analyzed bioinformatics of this sequence.The expression vector of pEGFP-N1-CTSZ was built and transfected into pig fibroblast cells with lipotectamine2000.The result was observation after 30 h.The result showed that the test successfully cloned the CDS sequence 912 bp of porcine CTSZ gene, and built pEGFP-N1-CTSZ fusion expression vector.The result of expression was green fluorescence in pig fibroblast cells by instantaneous transfection.This test laid a foundation for further study about the biology of this gene.%组织蛋白酶Z基因(Cathepsin Z gene, CTSZ)是影响猪生长、肉质等性状的一个重要候选基因.本研究以长白猪的背最长肌为试验材料,采用RT-PCR的方法克隆CTSZ基因CDS序列全长,对该序列进行了生物信息学分析,并构建pEGFP-N1-CTSZ融合表达载体,通过脂质体介导瞬时转染猪成纤维细胞,30 h后进行荧光表达检测.结果本试验克隆出猪CTSZ基因CDS全序列912 bp,并成功构建pEGFP-N1-CTSZ融合表达载体,瞬时转染后在猪成纤维细胞中表现为绿色荧光蛋白表达.本试验为进一步研究该基因的生物学功能奠定了基础.