柔嫩艾美耳球虫广东株GAM56蛋白的原核表达及其多克隆抗体的制备与鉴定

打开文本图片集

中图分类号： S852.72+3 文献标识码：A文章编码：1005-8567（2025）03-0001-07

Abstract：To elucidatethe biological functions of GAM56 protein inEimeria tenella，the coding sequence of the gam56 gene was amplified from the cDNA of gametocytes isolated from the Guangdong strain of E .tenella. Subsequently，a truncated prokaryotic expresion vector pET-B2M-GAM56 was constructed，and the recombinant GAM56 protein was expressed inE.coli expresionsystem using IPTG induction.Polyclonalantibodies were generated inBALB/c mice immunized with purified GAM56.The antibody titerand specificity were analyzed using indirect ELISA and Western blot. Gametocyte-stage samples of E .tenella（GD）were collected for immunofluorescence assay to evaluate antibody performance.Resultsrevealed that compared with the reference E .tenella（Houghton）strain，the gam56 gene of E . tenella（GD） exhibited a partial terminal deletion，presenting a 1O77bp CDS encoding 358 amino acids（predicted molecular weight 41.3 kDa）.The truncated GAM56 protein containing conserved regions was predominantly expressed ininclusion bodies.Indirect immunofluorescence assaydemonstrated specificrecognitionof theGAM56proteinatthe gametocytestage，with subcellularlocalizationconsistentwith literaturereports.This study provides critical materials for elucidating the biological functions of GAM56 in the E .tenella（GD）and establishes a foundation for functional investigations of sexual development-associated proteins in Eimeria species.

Keywords：Eimeria tenella；GAM56;Prokaryotic expression；Polyclonal antibodies

鸡球虫病是由一种或多种艾美耳球虫引起的严重危害家禽肠道健康的寄生性原虫病，给家禽养殖业造成重大损失[1-2]。（剩余8428字）