牛SRY基因的克隆与测序

Cloning and dequencing of SRY gene of bovine

以牛基因组DNA为模板,在一对特异性引物的引导下,利用PCR技术成功地扩增了牛SRY基因。纯化回收目的片段,克隆到pMD 18-T载体上,筛选阳性克隆测序,得到SRY基因两端的序列(5′端641 bp,3′端786bp)。与网上公布的牛SRY基因序列进行同源性比较,结果显示,牛SRY基因长度为2 713 bp,克隆的5′和3′端2个片段与网上公布的牛SRY基因序列对应片断的同源性分别为98.7%和98.1%。

The SRY gene was amplified by the polymerase chain reaction（PCR）with a pair of specific primers,then a 2 713 bp bovine SRY gene fragment with genomic DNA as the template was gained. The gene was purified and recovered and cloned into the pMD18-T vector,and the identified positive clone was sequenced to obtain 5＇and 3＇sequence （5＇end was 641 bp and 3＇end was 786 bp）. Compared with the SRY gene sequence on web,the result indicated that the SRY gene was 2 713 bp. The homologs of the two sequenced results and the bulletin result of bovine SRY gene are 98.7% and 98.1% ,respectively.