Epstein－Barr病毒潜伏膜蛋白（LMP）基因在哺乳动物传代细胞中的表达

EXPRESSION OF LATENT MEMBRANE PROTEIN(LMP)GENE OF EB VIRUS IN BABY HAMSTER DIDNEY CELL LINE

ＥＢ病毒潜伏膜蛋白（ＬＭＰ）是由病毒编码的主要的与病毒致宿主细胞潜伏感染有关的蛋白之一。我们用基因重组技术，把含有ＬＭＰ基因（ＢＮＬＦ１）３个外显子（ｅｘｏｎ）开放阅读框架（ＯＲＦ）的长１．８０ｋｂｐ的ＤＮＡ片段，和能分解ＨｙｇｒｏｍｙｃｉｎＢ的含有ＳＶ４０早期启动子和ＨｇｒｙｏｍｙｃｉｎＢ磷酸转移酶全基因（长１０２５ｂｐ）的ＤＮＡ片段（长１．６０ｂｐ），同时重组于亚克隆载体ｐＢｌｕｅｓｃｒｉｐｔＳＫ（ｐＢＳ）中，并使该重组质粒ｐＢＳ－ＬＭＰ－Ｈｙｇ（长５７６７ｂｐ）在乳地鼠肾传代细胞（ＢＨＫ）中获得表达。ＢＨＫ细胞在经此重组质粒转染后，ＬＭＰ阳性细胞是２％，在ＨｙｇｒｏｍｙｃｉｎＢ的持续压力下，ＬＭＰ表达细胞率可达２０％。３个月后，ＬＭＰ表达细胞逐渐减少。５个月后，不能测到ＬＭＰ表达细胞。经免疫荧光和蛋白印迹（Ｗｅｓｔｅｍｂｌｏｔ）实验证实，人鼻咽癌、风湿性关节炎和正常人血清中不含有抗ＬＭＰ抗体。

he recombinant plasmid pBluescript SK(pBS)-LMP-Hygwas successfully constructed.It encoded both the LMP and Hygromycin B phosphotransferase genes which expressed well in BHKcells.Cells transfected with recombinant plasmid pBS-LMP-Hyg are resistant to Hygromycin B(300μg/ml).The highest percentage of LMP positive cells is about 20%in BHK cells.TheLMP molecular weight is abut 60kDa,as detected by mouse anti-LMP monoclonalantibody by Western bolt.No anti-LMP antibody was detected in the sera from patients withnasopharyngeal carcinoma, patients with rheumatoid arthritis or normal individuals.