中国草食动物科学2026，Vol.46Issue(1)：58-64,7.DOI:10.3969/j.issn.2095-3887.2025.00.033
羊生长激素原核表达及抗血清的制备
Prokaryotic Expression and Antiserum Preparation of Ovine Growth Hormone
摘要
Abstract
This study aimed to utilize a prokaryotic expression system to express ovine growth hormone(oGH)protein,optimize induction conditions for protein purification,and obtain antiserum by immunizing New Zealand rabbits.The oGH gene was amplified using PCR and cloned into the pCold-MBP vector to construct the recombinant plasmid pCold-MBP-oGH.The recombinant plasmid was transformed into Escherichia coli BL21(DE3),and IPTG was added for induction to explore the effects of different IPTG induction conditions on the expression level of the fusion protein.The fusion protein was purified using a His-tagged nickel ion protein purification column,and Western blot was used to identify the fusion protein.The purified MBP-oGH fusion protein was mixed with Freund's adjuvant as an antigen to immunize two New Zealand rabbits,and antiserum was obtained after five immunizations.The results showed that the recombinant plasmid pCold-MBP-oGH was successfully constructed,and the MBP-oGH fusion protein was success-fully expressed.The optimal conditions for high expression of MBP-oGH were found to be:1.2 mM IPTG,37℃,for 5 hours.SDS-PAGE analysis identified the nickel column purification product as a single electrophoretic band of the tar-get protein,and Western blot results indicated that the pCold-MBP-oGH fusion protein could specifically bind to the corresponding antibody,with the antiserum from the two rabbits showing immunological specificity to the MBP-oGH fusion protein and displayed a single band.This experiment successfully constructed the recombinant plasmid pCold-MBP-oGH using a prokaryotic expression system,expressed and purified the MBP-oGH fusion protein,and immu-nized rabbits with the purified MBP-oGH fusion protein as an antigen,resulting in antiserum that exhibited immuno-logical specificity to the fusion protein.关键词
羊/生长激素/原核表达/蛋白纯化/抗血清Key words
ovine/growth hormone/prokaryotic expression/protein purification/antiserum分类
农业科技引用本文复制引用
阿迪莱·艾力,雷艳,赵树林,付强,薛慧婷,向志宇,赵红琼..羊生长激素原核表达及抗血清的制备[J].中国草食动物科学,2026,46(1):58-64,7.基金项目
国家自然科学基金(32060784) （32060784）
