牛妊娠相关糖蛋白10的制备及其生物信息学分析  

Preparation and bioinformatics analysis of pregnancy-associated glycoprotein 10 in cows

作　　者：王凯婷 陈柏桥 刘磊[2] 王会岩[1,2] WANG Kaiting;CHEN Baiqiao;LIU Lei;WANG Huiyan(Jilin Agricultural University,Changchun 130021,China;Jilin Collaborative Innovation Center for AntibodyEngineering,Jilin Medical University,Jilin 132013,China)

机构地区：[1]吉林农业大学生命科学院,吉林长春130021 [2]吉林医药学院吉林省抗体工程科技协同创新中心,吉林吉林132013

出　　处：《沈阳药科大学学报》2022年第8期993-1000,共8页Journal of Shenyang Pharmaceutical University

基　　金：吉林省科技厅技术攻关项目(20190301088NY)。

摘　　要：目的重组表达牛妊娠相关糖蛋白10(bovin pregnancy associated glycoproteins 10,BoPAG10),并对其结构和功能进行预测分析。方法通过密码子优化并合成BoPAG10基因,构建pcDNA3.1-BoPAG10重组表达载体,并瞬转至293F细胞中,采用Ni-NTA亲和层析对表达产物进行分离纯化,通过SDS-PAGE和Western blot检测表达效果,利用生物信息学软件对BoPAG10基因编码蛋白进行结构和功能预测。结果PCR和双酶切鉴定BoPAG10基因为1173 bp,成功表达BoPAG10重组蛋白,大小为48 kDa的,含量质量分数为91%。BoPAG10蛋白N端29个氨基酸为信号肽,7个糖基化位点和11个B细胞抗原表位。结论本研究作者成功表达并纯化出BoPAG10蛋白,为BoPAG10蛋白的结构和功能的研究以及抗BoPAG10单克隆抗体的制备奠定基础。Objective To obtain the recombinant bovine pregnancy associated glycoproteins 10(BoPAG10),and predict its structure and function.Methods Through codon optimization and synthesis of the BoPAG10 gene, the pcDNA3.1-BoPAG10 recombinant expression vector was constructed and transferred to 293 F cells.The expression product was separated and purified by Ni-NTA affinity chromatography.The expression effect was detected by SDS-PAGE and Western blot, and the structure and function of BoPAG10 gene-encoded protein were predicted by bioinformatics software.Results PCR and double enzyme digestion identified the BoPAG10 gene as 1 173 bp, and successfully expressed the BoPAG10 recombinant protein with a size of 48 kDa and a purity of 91%.The 29 amino acids at the N-terminal of BoPAG10 protein are signal peptides, 7 glycosylation sites and 11 B cell epitopes.Conclusion This study successfully expressed and purified BoPAG10 protein, laying a foundation for the study of the structure and function of BoPAG10 protein and the preparation of anti-BoPAG10 monoclonal antibodies.

关 键 词：牛妊娠相关糖蛋白 真核表达 纯化 生物信息学分析 

分 类 号：R917[医药卫生—药物分析学]