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目的 :获得单纯疱疹病毒 (HSV)新糖蛋白 g30K的部分特征氨基酸信息 ,以期对该蛋白基因的准确定位有所帮助。方法 :将针对HSV新糖蛋白 g30K的单克隆抗体 (mAb)CHA9生物素化后 ,淘筛噬菌体随机 12肽库 ,经 3轮筛选后进行ELISA检测。随机挑取 10个阳性克隆进行DNA测序 ,并进行序列比较以得到保守序列 ,然后对其疏水性进行预测。结果 :得到的保守氨基酸序列 (motif)为 PH/KHXHXGS 。携有此短肽的噬菌体可与CHA9特异结合而不与其它IgG出现交叉反应。疏水性分析证明其可能构成一个表位。结论 :筛选到一段具有含mAbCHA9靶抗原 HSV新糖蛋白 g30K部分氨基酸特征的短肽 ,为新基因的预测提供了参考依据 ,对进一步分析这一新糖蛋白的生物学功能也具有重要意义
Objective: To obtain partial amino acid information of HSV glycoprotein g30K in order to help the accurate localization of the protein gene. Methods: Monoclonal antibody (mAb) CHA9 against HSV neoglycoprotein g30K was biotinylated and screened for a random 12 - peptide phage library. After 3 rounds of screening, ELISA was performed. Ten positive clones were randomly selected for DNA sequencing and sequence comparison to obtain conservative sequences, and then their hydrophobicity was predicted. Results: The resulting conserved amino acid motif is PH / KHXHXGS. The phage carrying this short peptide can specifically bind to CHA9 without cross-reacting with other IgGs. Hydrophobicity analysis proved that it may constitute an epitope. CONCLUSION: A short peptide with the amino acid sequence of g30K containing the new glycoprotein of HSV containing mAbCHA9 target antigen was screened, which provided a reference for the prediction of new genes and was of great significance for further analysis of the biological function of this new glycoprotein