人巨细胞病毒p52蛋白基因的克隆及表达

CLONING AND EXPRESSION OF p52 GENE OF HUMAN CYTOMEGALOVIRUS

目的制备人巨细胞病毒（ＨＣＭＶ）ｐ５２蛋白特异性抗原。②方法用ＰＣＲ技术扩增ＨＣＭＶｐ５２蛋白ＩｇＭ抗原决定簇编码区ＤＮＡ片段，克隆到表达载体ｐＢＶ２２０中，转化Ｅ．ｃｏｌｉＤＨ５α株，用氨苄青霉素抗性和质粒酶切图谱分析法筛选阳性克隆，经温控诱导其表达。③结果重组克隆能有效表达天然蛋白ｐ５２，ＥＬＩＳＡ检测重组ｐ５２蛋白具良好的抗原特异性。④结论通过基因工程制备的重组ｐ５２蛋白可作为ＨＣＭＶ血清学诊断的良好抗原。

Objective To prepare p52 protein of human cytomegalovirus by using gene engineering technique. MethodsThe fragment encoding p52 epitope of IgM was amplified by PCR technique. The amplified PCR product was inserted into the expression vect or pBV220 and in turn tranformed strain E.coli DH5α. The recombinant clone was screened by ampicillin resistance and endonuclease digestion map of plasmid. The clone was induced by temperature variation. Results The recombinant clone could effectively express p52 nature protein. ELISA showed that the recombinant p52 had good specific antigenic characterization.Conclusion The recombinat p52 prepared by gene engineering technique is a good antigen in serum diagnosis of HCMV.