川西高原黑唇鼠兔乳酸脱氢酶C基因cDNA的克隆及序列分析

Cloning and Sequence Analysis of Lactate Dehydrogenase C (LDH-C) Gene from Black-lipped Pika in Western Sichuan Plateau

[目的]为研究以鼠兔LDH-C4为靶蛋白的节育型鼠药奠定基础。[方法]采用简并引物PCR克隆得到黑唇鼠兔LDH-C基因的EST序列,再根据EST序列采用RACE技术克隆出基因的开放阅读框(ORF)全长,和3’UTR序列。[结果]整个cDNA全长1 498 bp,其中ORF长996 bp,编码332个氨基酸,3’UTR长486 bp。ORF序列比对显示LDH-C基因在大的分类单位上均很保守。系统树分析显示黑唇鼠兔的LDH-C基因和灵长类及偶蹄目的遗传距离较近,和啮齿目较远。[结论]成功的克隆得到了黑唇鼠兔的LDH-C基因的cDNA序列。

[Objective] The aim was to lay a foundation for studying contraceptive rodenticide with LDH-C4 as target protein.[Method] EST sequence of LDH-C gene from black-lipped pika was cloned by PCR with degenerate primers;then the full length of open reading frame（ORF） and 3′UTR sequence were cloned by RACE technique.[Result] The full length of cDNA was 1 498 bp containing an ORF of 996 bp and a 3′UTR of 486 bp.The ORF encoded a polypeptide of 332 amino acids.The alignment of LDH-C gene ORF nucleotide sequences from different species showed that the gene was conserved even between large taxons.The phylogenic tree showed that black-lipped pika LDH-C was closer to primates and artiodactyla than to rodents.[Conclusion] cDNA sequence of LDH-C gene from black-lipped pika was cloned successfully.