文献类型：期刊文献

英文题名：Prokaryotic expression, polyclonal antibody production, and application of yak TGF-β2

作者：Chen, Yaming[1,2];Pan, Yangyang[1,3];Yu, Sijiu[1,3];Wang, Jinglei[1,3];Fan, Jiangfeng[1,3]

第一作者：陈亚明;Chen, Yaming

通信作者：Fan, JF[1];Fan, JF[2]

机构：[1]Gansu Agr Univ, Coll Vet Med, Lanzhou, Gansu, Peoples R China;[2]Gansu Univ Chinese Med, Sch Clin Med 1, Lanzhou, Gansu, Peoples R China;[3]Technol & Res Ctr Gansu, Lanzhou, Gansu, Peoples R China

第一机构：Gansu Agr Univ, Coll Vet Med, Lanzhou, Gansu, Peoples R China

通信机构：[1]corresponding author), Gansu Agr Univ, Coll Vet Med, Lanzhou, Gansu, Peoples R China;[2]corresponding author), Technol & Res Ctr Gansu, Lanzhou, Gansu, Peoples R China.

年份：2025

卷号：22

期号：4

外文期刊名：ANIMAL REPRODUCTION

收录：;Scopus(收录号：2-s2.0-105019657566);WOS:【SCI-EXPANDED(收录号:WOS:001606324800002)】；

基金：This work was supported by the National Natural Science Foundation of China (Nos. 31972760, 32160859) . Outstanding Youth Foundation of Gansu Province (No. 20JR10RA561) and the Innovation and Entrepreneurship Capacity Building Project of the Gansu Provincial Department of Education (No. 2019B-081) . Fuxi Young Talents Fund of Gansu Agricultural University (No. Gaufx-02Y10) and the Scientific Research Startup Fund for Doctoral Scholars of Gansu Agricultural University (No. GSAU-RCZX201701) .

语种：英文

外文关键词：yak; TGF-beta 2; prokaryotic expression; polyclonal antibody

摘要：This study aimed to generate yak-specific polyclonal antibodies against transforming growth factor beta 2 (TGF-beta 2). Specific primers targeting the TGF-beta 2 coding sequence (CDS) were designed, and the gene was amplified via RT-PCR. The amplified product was cloned into the pET-32a(+) vector to construct the recombinant plasmid pET-32a(+)-TGF-beta 2. This plasmid was transformed into Escherichia coli BL21(DE3) for protein expression. Isopropyl beta-D-1-thiogalactopyranoside (IPTG) induced TGF-beta 2 production, and the recombinant protein was purified. New Zealand rabbits were immunized with the purified protein to generate polyclonal antibodies. Polyclonal antibody titers were determined using ELISA, while specificity was assessed through Western blot and immunohistochemistry. The recombinant plasmid was successfully constructed, and IPTG induction yielded a 63 kDa protein. Optimal expression occurred at 25 degrees C with 0.5 mmolL-1 IPTG and a 10-hour induction period. ELISA confirmed an antibody titer of 1:106. Western blot and immunohistochemistry demonstrated TGF-beta 2 expression in female yak ovaries, oviducts, and uteri across reproductive stages, with significantly elevated ovarian levels during pregnancy. This study successfully produced and validated a highly specific anti-yak TGF-beta 2 polyclonal antibody, providing a vital tool for investigating its role in yak reproductive physiology.