CAJ | 학술논문

将麻疹病毒(Nepal株)的血凝素(hemagglutinin)基因插入真核表达载体pIRES-EGFP,并在HeLa细胞中表达.因其较低的表达量,所以将其截短,去除跨膜区.使这个截短的HA基因与绿色荧光蛋白基因融合,并克隆至原核表达载体pET-28b中.将重组质粒转入大肠杆菌中表达,产生了分子量约为90kD的融合蛋白.通过ELISA和Western印迹来检测这个基因工程蛋白的抗原性.在检测一系列的血凝素阳性或阴性的人血清中,这个融合蛋白的阳性检出率为90%,阴性检出率为100%(与市售麻疹病毒诊断试剂盒相比较).由于此HA蛋白是原核表达产物,回避了真核表达系统复杂的操作过程和昂贵的费用,所以,这个麻疹病毒血凝素基因工程抗原有望成为一种新型、便捷的麻疹病毒诊断试剂.
장마진병독(Nepal주)적혈응소(hemagglutinin)기인삽입진핵표체재체pIRES-EGFP,병재HeLa세포중표체.인기교저적표체량,소이장기절단,거제과막구.사저개절단적HA기인여록색형광단백기인융합,병극륭지원핵표체재체pET-28b중.장중조질립전입대장간균중표체,산생료분자량약위90kD적융합단백.통과ELISA화Western인적래검측저개기인공정단백적항원성.재검측일계렬적혈응소양성혹음성적인혈청중,저개융합단백적양성검출솔위90%,음성검출솔위100%(여시수마진병독진단시제합상비교).유우차HA단백시원핵표체산물,회피료진핵표체계통복잡적조작과정화앙귀적비용,소이,저개마진병독혈응소기인공정항원유망성위일충신형、편첩적마진병독진단시제.
Hemagglutinin (HA)gene of measles virus (MV)( Nepal strain)was cloned into eukaryotic expression vectorpIRES-EGFP, and the recombinant plasmid was transfected into HeLa cells. Beacause of the low expression of ha gene in HeLa cells, ha gene was truncated and cloned into prokaryotic expression vector pET-28b which contained egfo gene.Then, the recombinant plasmid was transformed into E. Coli BL21 DE3, producing a fusion antigenic protein H/EGFP with molecular weight of 90 kD. The engineered HA protein was assayed by using ELISA and Western-blotting for detection the antibodies to measles virus in a cohort of hemagglutinin-positive of -negative human sera. The results showed that the MV hemagglutinin gene engineering antigen might provide a more efficient diagnostic reagent for detection of the MV infection