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The cDNA was cloned into the bacterial expression vector pET28a(+) and expressed in<jats:italic>E. coli<\/jats:italic>BL21 cells. The antigenicity of the recombinant gametocyte protein and its localization in different<jats:italic>E. necatrix<\/jats:italic>life-cycle stages were determined by western blot and indirect immunofluorescence analyses, respectively.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>A 731-nucleotide sequence of cDNA [GenBank: KF649255] of<jats:italic>E. necatrix<\/jats:italic>had 97.7% identity to that of Et<jats:italic>gam22<\/jats:italic>of<jats:italic>E. tenella<\/jats:italic>. The cDNA ORF encoded a 186-amino acid protein containing a histidine-proline-rich region. The recombinant gametocyte protein (rEnGAM22) was predominately expressed in the insoluble inclusion body and recognized by antiserum from chickens immunized with oocysts of<jats:italic>E. necatrix<\/jats:italic>,<jats:italic>E. maxima<\/jats:italic>and<jats:italic>E. tenella<\/jats:italic>. A specific antibody to the rEnGAM22 protein recognized the wall-forming bodies in macrogametocytes and the walls of oocysts and sporocysts.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>The gene cloned from<jats:italic>E. necatrix<\/jats:italic>gametocytes is an ortholog to Et<jats:italic>gam22<\/jats:italic>of<jats:italic>E. tenella<\/jats:italic>and presents a potential target for future recombinant subunit vaccines against coccidiosis.<\/jats:p><\/jats:sec>","DOI":"10.1186\/1756-3305-7-27","type":"journal-article","created":{"date-parts":[[2014,1,15]],"date-time":"2014-01-15T16:24:59Z","timestamp":1389803099000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":39,"title":["Cloning and characterization of an Eimeria necatrix gene encoding a gametocyte protein and associated with oocyst wall formation"],"prefix":"10.1186","volume":"7","author":[{"given":"Dandan","family":"Liu","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Liqin","family":"Cao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yulan","family":"Zhu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Changjing","family":"Deng","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shijie","family":"Su","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jinjun","family":"Xu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wenjie","family":"Jin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jingui","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lili","family":"Wu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jianping","family":"Tao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2014,1,15]]},"reference":[{"issue":"1","key":"1174_CR1","doi-asserted-by":"publisher","first-page":"58","DOI":"10.1128\/CMR.15.1.58-65.2002","volume":"15","author":"PC Allen","year":"2002","unstructured":"Allen PC, Fetterer RH: Recent advances in biology and immunobiology of Eimeria species and in diagnosis and control of infection with these coccidian parasites of poultry. 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