猪唾液酸粘附素的分段表达及功能鉴定
Expression and characterization of PRRSV sialoadhesin fragments
为了验证猪pSn与PRRSV囊膜糖蛋白GP5的关系，提取PAM总RNA，用分段RT-PCR方法克隆得到猪pSn全长基因，并进行了核苷酸序列测定。将5个分段基因克隆入pET-28a，进行诱导表达和纯化，用SDS-PAGE和Western blot方法对表达的蛋白进行了鉴定，用ELISA方法检测了5个分段蛋白与分段表达的GST融合GP5分段蛋白的相互关系进行了研究。结果表明扩增出了pSn的5个分段基因，与GenBank中登录的序列的同源性为99%。5个分段基因克隆人pET-28a，经诱导后用SDS-PAGE和Western blot验证均得到了表达。ELISA结果表明PSn-3能与GST-GP5-1蛋白结合。通过用ELISA方法鉴定分段表达pPSn蛋白片段与分段表达的GP5的关系，表明pPSn-3能与GST-GP5-1蛋白结合，为了解PRRSV的感染细胞机制奠定基础。
In this paper to identify the relationship of sialoadhesin and envelope glycoprotein GP5, 5 fragments of whole procine Sn (pSn) were cloned from porcine alveolar phagocyte cells by RT-PCR. After all the fragments sequenced they were cloned into pET-28a vector and expressed. The purified proteins were identified by SDS-PAGE and Western blot. The relationship of fragments proteins and fragments of GST-GP5 were analysed by ELISA. The results showed that all the fragments were cloned and expressd. They all were the target proteins by identification of SDS-PAGE and Western blot. The results of ELISA showed that fragment of PPSn-3 could bind with GST-GP5-1. The binding of pSn with GP5 was confirmed by identification of ELISA, and it would help to learn more about the infection mechanism of PRRSV.
邱洪凯、崔然、肖一红、刘岳、周恩民
分子生物学生物工程学
猪繁殖与呼吸综合征病毒唾液酸粘附素GP5表达功能
PRRSVsialoadhesinGP5expressionfunction
邱洪凯,崔然,肖一红,刘岳,周恩民.猪唾液酸粘附素的分段表达及功能鉴定[EB/OL].(2011-03-31)[2025-07-16].http://www.paper.edu.cn/releasepaper/content/201103-1190.点此复制
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