奶牛妊娠相关糖蛋白2的表达及结构功能预测分析

doi:10.16431/j.cnki.1671-7236.2020.10.001

Abstract

Abstract: In order to obtain bovine pregnancy-associated glycoprotein 2 (BoPAG2) with high expression and purification and good antigenicity,the structure and function of the protein were analyzed by bioinformatics prediction.The BoPAG2 gene of Holstein dairy cow was amplified by PCR,and the gene was ligated into the expression vector pET-28a,transformed into E.coli (BL21) to induce expression and purification.SDS-PAGE and Western blotting were used to verify and detect the size and immunological characteristics of the protein.The glycosylation modification,B-cell epitope,hydrophilicity,signal peptide,secondary structure and 3D model,conservative domain and protein interaction of BoPAG2 protein were predicted and analyzed by bioinformatics.The results showed that the BoPAG2 gene was successfully cloned,and the corresponding protein was expressed and purified by induction.The size of the BoPAG2 protein was consistent with the expected results by SDS-PAGE and Western blotting.The results of bioinformatics prediction analysis showed that five sites of BoPAG2 protein occurred glycosylation modification,Asp⁵¹,Asp⁷¹,Asp¹¹⁴,Asp²⁵² and Asp³⁴³,respectively.The protein contained 15 B-cell epitopes and a signal peptide sequence consisting of 15 amino acids in the N terminal.The 3D model of the BoPAG2 protein was successfully established.The protein contained 4 superfamily domains.The results of protein interaction analysis showed that there were 7 proteins that interact with BoPAG2.The protein might be involved in digestion and absorption.Other processes in the maternal regulatory function during pregnancy.In summary,the BoPAG2 protein was successfully expressed and purified in this experiment.The bioinformatics prediction analysis of the structure and function of BoPAG2 protein provided a theoretical basis for the preparation and function research of BoPAG2 protein antibody.

Key words: pregnancy-associated glycoproteins; prokaryotic expression; antigen epitopes; signal peptide

CLC Number:

Q785

YANG Yajun, HE Jinke, BAI Tiange, SONG Shengnan, YANG Ningning, CHEN Chuangfu, WANG Zhen. Expression and Structure Function Prediction of Bovine Pregnancy-associated Glycoprotein 2[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(10): 3039-3048.

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Expression and Structure Function Prediction of Bovine Pregnancy-associated Glycoprotein 2

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