民猪CIRP基因的克隆与冷诱导研究

Cloning and cold induction of Min pig CIRP gene

采用生物信息学结合RT-PCR的方法,克隆民猪的冷诱导RNA结合蛋白(Cold inducible RNA-binding protein,CIRP)基因,获得3个变异体序列。猪CIRP变异体1基因cDNA长1 278 bp(GenBank登录号:HQ908794),编码172个氨基酸;变异体2基因cDNA长1 653 bp(GenBank登录号:HQ908795),编码182个氨基酸;变异体3基因cDNA长1 765 bp(Genbank登录号:HQ908796),编码144个氨基酸。CIRP变异体2与变异体1相比,在其编码区的第501碱基处,出现了一段375 bp的插入片段,该插入片段的第46～48个碱基为TAG,使翻译提前终止;变异体3与变异体2相比,在其编码区的第428碱基处,出现一段115 bp插入片段,该插入片段的第5～7个碱基为TGA,使翻译提前终止。3种转录变异体的核苷酸序列同源性为86.45%,氨基酸序列同源性为87%。冷诱导后,CIRP基因在肌肉中的表达水平出现显著升高(P<0.05)。

Min pig CIRP gene was cloned by bioinformatics, RT-PCR and three variant sequences were obtained. Variant 1 of Min pig CIRP is 1 278 bp in length （GenBank accession number： HQ908794）, and it encodes 172 amino acids. Variant 2 is 1 653 bp in length （GenBank accession number： HQ908795）, and it encodes 182 amino acids. Variant 3 is 1 765 bp in length （GenBank accession number： HQ908796）, and it encodes 144 amino acids. Variant 2 compared to variant 1, there was an insert fragment of 375 bp in its encoding region of the 501 base. The forty-sixth to the forty-eighth bases of the insert fragment were TAG, which lead to translation termination. Variant 3 compared to Variant2, there was an insert fragment of 115 bp in its encoding region of the 428 base. The fifth to the seventh were TGA, which lead to translation termination. Nucleic acid sequence homology of CIRP variant is 86.45%, and Amino acid sequence homology of CIRP variant is 87%. Cold induced C/PR mRNA expression level increased si.qnificantlv in muscle （P〈0.05）.