用点突变的方法组建巨细胞病毒抗原决定簇基因的表达克隆

研究资料表明，人巨细胞病毒（ＨＣＭｖ）单一蛋白的单一抗原决定簇只能被部分患者阳性血清识别。组建在血清学诊断中能够替代全病毒抗原的基因工程抗原，需要含有病毒多种主要抗原蛋白的抗原决定簇。为搞清在表达载体中重复插入某一抗原决定簇基因是否能表达出更高抗原效价的融会蛋白，我们用点突变的方法，在表达载体中分别插入了人ＨＣＭｖ的ｐｐＵＬ３２蛋白羧基端一个抗原决定簇基因的１个、２个和３个拷贝。在免疫转印检测中，这些克隆表达的融合蛋白与特异性阳性血清的反应性差别不明显。这表明，插入表达载体中目的基因的多寡对表达蛋白的抗原效价没有显著影响。

CONSTRUCTION OF A EXPRESSION CLONE OF AN EPITOPE GENE OF HUMAN CYTOMEGALOVIRUS BY MUTAGENESIS

he data have shown that single epitope of HCMV protein could only be recognized by partial positive serafrom patients.A gene clone antigen that can replace virion antigen in serological diagnosis need to be composed ofmultiple epitopes of several main antigenic proteins of the virus.To understand whether a clone containing repeatgene of epitope can express fusion protein with high antigenic titer,we have constructed 3 clones containing 1,2,and 3 copies of the epitope gene of HCMV ppUL32 C-terminal respectively by mutagenesis.There was no signifi-cant difference of the reactions between the fusion proteins expressed from the 3 clones and viral positive sera.Theresults showed that the different numbers of epitope genes in expression clones had no significant effect on the anti-genic titers of their fusion proteins.