猪Calsarcin-2基因编码区的克隆及组织表达谱分析

Cloning of Encoding Region and Analysis of Tissue Expression Profile of Pig Calsarcin-2 Gene

目的克隆猪Calsarcin-2(Cs-2)基因,并探讨其在猪组织中的表达特点。方法利用RT-PCR方法扩增猪Cs-2基因编码区的序列;采用Northern杂交和半定量RT-PCR方法在猪不同组织和不同部位骨骼肌中分析Cs-2基因表达情况及转录本个数、大小。结果猪Cs-2基因片段长934 bp,含有1个897 bp的开放阅读框(ORF),编码298个氨基酸,与已报到的人Cs-2基因氨基酸序列相似性为93%;猪Cs-2基因仅在骨骼肌中表达,不同骨骼肌中表达量有所不同,以背最长肌最高;此基因在骨骼肌中只有一个转录本,大小为1.26 kb。结论猪Cs-2基因序列与已报道的人、鼠Cs-2基因具有高度同源性,主要在骨骼肌中特异表达。

Objective To clone swine Cs -2 gene and to investigate the character of tissue expression in pigs. Methods The sequence of porcine gene Cs - 2 was amplified by RT - PCR. The numbers and size of porcine Cs - 2 transcripts in skeletal muscle were obtained with Northern blotting and the expression levels in multiple tissues were detected by semi - quantitative RT - PCR. Resuits The length of porcine Cs -2 cDNA was 934 bp containing an ORF of 897 bp, which encoded a 166 amino acids. Homology of amino acids sequence between the amplified porcine and the reported human Cs -2 was 93%. There was only one transcript among different skeletal muscles whose size was nearly 1.26 kb, but not detected in the other tissues including heart, liver, spleen, lung and kidney. The expression levels of Cs - 2 gene varied among different skeletal muscles. It was the highest expression in musculus longissimus dorsi. Conclusions There are great similarities in the porcine Cs -2 gene, human gene and murine Cs -2 gene, which are expressed mainly in skeletal muscles.