新疆部分地区牛轮状病毒VP6基因的原核表达与免疫反应性分析

Prokaryotic expression and immunoreactivity analysis of bovine rotavirus VP6 gene in parts of Xinjiang

为了原核表达牛轮状病毒VP6蛋白,检测该蛋白的反应原性,对牛轮状病毒内衣壳蛋白VP6保守区设计并合成引物,表达牛轮状病毒VP6蛋白,应用Western Blot分析该蛋白的反应原性。结果显示,利用PCR方法成功扩增出目的基因,经过测序与酶切鉴定,正确构建了表达载体pET-32a-VP6,SDS-PAGE电泳显示表达产物在60 k D表达重组蛋白。表明经过Western Blot检测,该蛋白具有良好的反应原性,为后续该蛋白作为诊断抗原奠定了基础。

Bovine rotavirus VP6 protein was expressed by prokayotic expression vector and its immune reactivity was analyzed.Primers were designed for the VP6 conserved region of the bovine rotavirus lingerie shell protein,and the immune reactivity of the expressed protein was analyzed by Western Blot. Results showed that the target gene was successfully amplified by RT-PCR method.After sequencing and enzyme digestion,the expression vector pET-32 a-VP6 was constructed correctly,and the recombinant protein was 60 Da by SDS-PAGE electrophoresis. The protein showed good reactivity by western blot dectection.