柔嫩艾美耳球蟲廣東株GAM56蛋白的原核表達及其多克隆抗體的制備與鑒定

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中圖分類號： S852.72+3 文獻標(biāo)識碼：A文章編碼：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

雞球蟲病是由一種或多種艾美耳球蟲引起的嚴(yán)重危害家禽腸道健康的寄生性原蟲病，給家禽養(yǎng)殖業(yè)造成重大損失[1-2]。（剩余8428字）