Prokaryotic Expression of the GAM56 Protein from Eimeria tenella Guangdong Strain and Preparation and Identification of Its Polyclonal Antibodies
JIANG Minghui
SONG Yongle
SU Guanzhi
CHEN Qingbiao
LI Juan
QI Nanshan
LIAO Shenquan
LV Minna
LIN Xuhui
CAI Haiming
ZHANG Xu
SUN Mingfei
Abstract：To elucidate the biological functions of GAM56 protein in Eimeria 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 expression vector pET-B2M-GAM56 was constructed,and the recombinant GAM56 protein was expressed in E.coli expression system using IPTG induction.Polyclonal antibodies were generated in BALB/c mice immunized with purified GAM56.The antibody titer and 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.Results revealed that compared with the reference E.tenella(Houghton)strain,the gam56 gene of E.tenella(GD)exhibited a partial terminal deletion,presenting a 1 077 bp CDS encoding 358 amino acids(predicted molecular weight 41.3 kDa).The truncated GAM56 protein containing conserved regions was predominantly expressed in inclusion bodies.Indirect immunofluorescence assay demonstrated specific recognition of the GAM56 protein at the gametocyte stage,with subcellular localization consistent with literature reports.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 tenellaGAM56Prokaryotic expressionPolyclonal antibodies
Publication Date：2025-06-18
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：7( 1-7 )