摘要
目的通过克隆广西巴马小型猪活化素(activin)βA/βB基因cDNA,分析其序列,为研究活化素的生物学功能打下基础。方法提取广西巴马小型猪发情母猪的卵巢总RNA,并以其为模板,以下游特异性引物为反转录引物,在AMV酶作用下合成cDNA;用LA-Taq进行PCR扩增出包括成熟肽在内的activinβA及βB基因的部分序列,把两个序列分别连接到PMD-18载体进行克隆和序列分析。结果包括成熟肽在内的activinβA及βB基因的部分序列长度分别为964 bp和791 bp,两个亚基的序列比较保守,与GenBank上发表的猪activin两个亚基同源性达到99.79%和99.62%。activinβA基因有两个碱基发生突变,其中一个在成熟肽序列上,一个在非编码序列上;βB基因信号肽序列上有三个碱基发生突变,其中一个是同义突变。
In this study, total mRNA isolated from the ovary tissue of Guangxi Bama mini-pig were used as templates to synthesize cDNA by reverse transcription polymerase AMV. The partial sequence of activin βA and βB cDNA with length of 964 bp and 791 bp respectively, was obtained by PCR. The homology analysis showed that they are highly conservative. The homology rate was 99.79 % and 99.62 % respectively, compared with the activin βA and βB gene of other pig breeds reported in GenBank. Two nucleotides mutation were found in activin βA gene, of which one mutation occurred in βA mature peptide and the other in none coding part. Three nucleotides mutations occurred in the signal peptide of βB, of which one is silent mutation.
出处
《实验动物科学》
2007年第6期1-5,共5页
Laboratory Animal Science
基金
国家科技攻关计划(2004BA717B-01-03)
广西科学研究与技术开发计划项目(桂科能0630006-6B)
广西自然科学基金项目(桂科自0542025)
