山羊SDHD基因cDNA编码序列的克隆与分析

利用NCBI中GenBank里查询到已登录的人、猪、牛和绵羊的SDHD mRNA序列,通过多重同源比较,从而获得高度保守区域序列,设计了同源引物,并首次对山羊SDHD编码序列进行了分子克隆。经过PCR扩增和测序,获得山羊SDHD基因共4段cDNA序列,分别为:451 bp4、20 bp5、29 bp和698 bp。所获得的四段序列测序结果经La-sergene7.0软件SeqMan拼接后,获得一条1238 bp长的cDNA序列。在GenBank数据库中进行BLAST/nr比对,发现其与绵羊、牛、猪和人相应序列相似性分别达98%、97%、85%和81%。用NCBI的ORF Finder软件对已经克隆的山羊SDHD基因cDNA进行开放阅读框分析,发现该序列包含一个480 bp的开放阅读框,编码159个氨基酸残基,计算机分析表明(Compute pI/Mw tool),该蛋白的分子量约为17 224.11 Da,等电点为8.92。通过DNAMAN软件分析发现,山羊SDHD蛋白保守性很高,其与绵羊、牛、猪和人SDHD蛋白在氨基酸序列上的相似性分别达到97%、96%、87%和85%。此山羊SDHD基因cDNA序列和SDHD蛋白质序列已于2010年1月31日登录在NCBI的GenBank上,登录号为GU338978和ADB92501。本项研究为进一步研究山羊的SDHD基因作为山羊肉品质性状候选基因,提供了相应的序列信息。

Cloning and Bioinformatic Analysis of cDNA Encoding Caprine SDHD Gene

The present study is using the accessed sequences of human,pig,cattle and sheep SDHD mRNA that checked from GenBank of NCBI,through multiple alignment,high conservative sequences were found,and homologized primers were designed based on them.This is the first time to clone SDHD gene from goat genome.By PCR amplification and sequencing,four cDNA sequences of goat SDHD gene are gotton,they are 451 bp,420 bp,529 bp and 698 bp in length,respectively.Through splicing these four cDNA sequences by SeqMan software of Lasergene,getting a 1238 bp length of cDNA sequence.The similarity of this sequence is up to 98%,97%,85% and 81% respectively aligned with corresponding mRNA sequences of sheep,cattle,pig and human in BLAST/nr of GenBank database.Analyzing this sequence by ORF Finder software,it can find out a 480 bp open reading frame,coding 159 amino acids.Then analyzing this amino acid sequence by Compute pI/Mw tool,it is computed out that this protein molecular weight is about 17 224.11 Da,and isoelectric point is about 8.92.Analyzing this protein sequence by DNAMAN software,the similarity is up to 97%,96%,87% and 85% respectively compared with corresponding protein sequences of sheep,cattle,pig and human.This caprine SDHD gene sequence and protein sequence had been submitted to GenBank of NCBI at 31-JAN-2010.The GenBank Accession number is GU338978 and ADB92501,respectively.Our present sdudy will provide sequence information of caprine SDHD gene for further studying as a candidate gene of meat quality trait in goats.