摘要从天祝白牦牛的基因组DNA中扩增了SRY（Sex-determining Region on the Y Chromosome，SRY）基因编码区序列，将其克隆至pGEM-T easy载体并送至生物公司测序，天祝白牦牛SRY基因编码区长687 bp，编码229个氨基酸；对牦牛和奶牛的SRY基因编码区进行序列比对，发现存在2个碱基的变异，造成1个氨基酸的变异；将牦牛SRY基因编码区连接至pET-28a(+)载体，成功构建了表达载体pET-28a/SRY；把表达载体pET-28a/SRY转入大肠杆菌E.coli BL21（DE3）中，在合适的条件下诱导该大肠杆菌，SRY蛋白得到了大量表达；对表达产物进行了Western-blot检测，进一步确定牦牛SRY蛋白得到表达。
Abstract：The Tianzhu White Yak’s SRY gene was amplified from Yak’s genome. The product was cloned to pGEM-T easy vector and sequenced by biological company. The total coding region of Tianzhu White Yak’s SRY gene is 687 bp, encoding a peptide with 229 amino acid residues. Through sequences aligned, it was discovered that there are 2 mutations in the coding region between yak and cow, causing 1 amino acid mutation in LF protein. The coding region of Yak’s SRY gene was cloned to EcoR I and Sal I sites of pET-28a(+) vector to construct an expression plasmid pET-28a/SRY. The expression plasmid was transformed to E.Coli BL21 (DE3), and induced under proper conditions then the SRY protein was highly expressed. The product of expression was identified by Western-bloting to be sure that the SRY protein of yak was expressed.
