中文摘要：

为确定人巨细胞病毒M抗原表位MAD的关键氨基酸残基，以MAD多肽序列为基础，分别将保守氨基酸残基单一突变为甘氨酸残基，构建各自突变体，然后与人源Fc的N端融合，通过原核表达载体pET32-Fc表达融合蛋白MAD-Fc，经proteinA柱亲和纯化得到各突变体纯品。通过ELISA及Westernblotting方法验证各突变体特异结合羊抗HCMV多抗间的差异，从而确定表位关键氨基酸残基。结果显示，将MAD中的谷氨酰胺残基单突变为甘氨酸残基后，MADo—G结合羊抗HCMV多抗活性大大降低，差异显著；而其他氨基酸残基单突变时，对MAD活性影响程度很小。由此得出结论：MAD结合羊抗HCMV多抗的活性与谷氨酰胺残基有关。

英文摘要：

We identified the critical amino-acid residues in antigen M derterminant（MAD） epitope of human cytomegalovirns protein M. On the basis of the peptide sequence of MAD, some conservative residues were mutated into the glycine residue. Then the gene fragment of mutants linked to amino terminal of Fc were cloned into the plasmid pET32-Fc and expressed by fusion with Fc. After purified by protein A affinity chromatography, the activity of mutants binding the goat polyclonal antibodies against human cytomegalovirns（HCMV） were detected by ELISA and Western blotting. Our results showed that when glutamine residue was mutated into glycine residue, the activity of MADQ→G binding the goat polyclonal antibodies against HCMV was reduced apparently. Other mutants did not have the same characteristics. The activity of MAD was closely related to the conformation of glutamine residue.