水牛SRY基因的克隆及序列分析

根据荷斯坦牛SRY基因设计一对引物，采用聚合酶链式反应（PCR）技术，以中国沼泽型水牛（Swamp Buffalo）基因组DNA为模板，扩增得到SRY（Sex Deterimation region of Y chromosome）全序列约2005bp，其中1-504bp为5’启动子区，1196-2005bp为3’侧翼序列，在505-1195bp为SRY的外显子，编码229个氨基酸。在SRY HMG box区域设计探针，用地高辛标记后分别与雄性、雌性水牛基因组DNA进行Southern 杂交，结果显示该段序列只在雄性DNA样本中有杂交信号，证明SRY基因为雄性特异。BLAST比对结果显示与牛属动物SRY基因的同源性为96％，其中SRY基因HMG box区域同源性高达99%，说明SRY基因具有高度的进化保守性

Cloning and characterization of swamp Buffalo SRY gene

The SRY gene from buffalo (Bubalus bubalis) genome was amplified by the polymerase chain reaction ( PCR) with primers based on the sequence of Hostein SRY gene. The amplified fragment was 2005 bp include 5UTR ( 1 - 504bp) and 3'UTR(1196 - 2005bp). And the amplified fragment was cloned and sequenced. Sequence analysis showed that the coding region of SRY gene (505 - 1195bp) from buffalo was highly homologous with those of other bovine counterpart genes (96% homology) , especially in the HMG box region (99%homology). It was found that there were only signal on male buffalo genome on Southern blot,which indicate SRY gene are highly conservative on evolves.