人细小病毒B19 NS1片段全长的克隆及表达

目的 :克隆人细小病毒B1 9非结构蛋白 1 (NS1 )全长基因 ,构建重组表达载体并诱导表达。　方法 :利用聚合酶链反应 (PCR)和分子克隆技术 ,从我科保存的B1 9阳性标本XA2 0中扩增NS1片段全长 ,构建NS1 pGEM T easy克隆载体并测序分析 ,测序证实序列正确后亚克隆于pQE30表达载体中 ,并转化至M1 5感受态菌中 ,经IPTG诱导可表达 770 0 0的融合蛋白。　结果 :成功克隆了人细小病毒B1 9NS1全长基因 ,构建了融合蛋白NS1 pQE30的重组表达质粒 ,表达的融合蛋白经 1 2 %SDS PAGE电泳证实为 770 0 0。　结论 :获得人细小病毒B1 9NS1全长基因及原核表达产物 ,对进一步研究NS1的生物学活性及其单克隆抗体的制备有重要意义。

The clone and express of NS1 of human parvovirus B19

Objectives:To clone human parvovirus B19 NS1 gene, construct the recombinant vector, express its product. Methods:NS1 gene was amplified by PCR from Xi'an 20 strain kept by our laborary and cloned into pGEM T easy. After sequencing, it was subcloned into pQE30 plasmid. After 4h induced by IPTG,the expression of recombinant Mr 77 000 fusion protein NS1 pQE30 was confirmed by SDS PAGE. Results:Human parvovirus B19 NS1 gene was coloned. Recombinant expression plasmid NS1 pQE30 was constructed. The expressed fussion protein was confirmed by SDS PAGE. Conclusions:The human parvovirus B19 NS1 gene and prokaryotic expression products were obtained. It might be important for study on the function of human parvovirus B19 NS1 and preparing the monoclonal antibody against human parvovirus B19 NS1.