Cloning and characterization of an Eimeria necatrix gene encoding a gametocyte protein and associated with oocyst wall formation
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- 作者：Dandan Liu (10)
Liqin Cao (10)
Yulan Zhu (10)
Changjing Deng (10)
Shijie Su (10)
Jinjun Xu (10)
Wenjie Jin (10)
Jingui Li (10)
Lili Wu (10)
Jianping Tao (10) - 关键词：Eimeria ; Gametocyte protein ; Gene ; Cloning and expression ; Immunolocalization
- 刊名：Parasites & Vectors
- 出版年：2014
- 出版时间：December 2014
- 年：2014
- 卷：7
- 期：1
- 全文大小：857 KB
- 作者单位：Dandan Liu (10)
Liqin Cao (10)
Yulan Zhu (10)
Changjing Deng (10)
Shijie Su (10)
Jinjun Xu (10)
Wenjie Jin (10)
Jingui Li (10)
Lili Wu (10)
Jianping Tao (10)
10. Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Ministry of Education Key Lab for Avian Preventive Medicine, Key Lab of Jiangsu Preventive Veterinary Medicine, College of Veterinary Medicine, Yangzhou University, 12 East Wenhui Road, Yangzhou, Jiangsu, 225009, People鈥檚 Republic of China - ISSN：1756-3305
文摘
Background Gametocyte proteins of Eimeria (E.) spp. are important components of the oocyst wall and some have been used to develop transmission-blocking vaccines against avian coccidiosis. Methods Total RNA isolated from E. necatrix gametocytes was utilized as templates for RT-PCR amplification and sequencing of cDNA encoding a gametocyte protein using gene-specific primers. The cDNA was cloned into the bacterial expression vector pET28a(+) and expressed in E. coli BL21 cells. The antigenicity of the recombinant gametocyte protein and its localization in different E. necatrix life-cycle stages were determined by western blot and indirect immunofluorescence analyses, respectively. Results A 731-nucleotide sequence of cDNA [GenBank: KF649255] of E. necatrix had 97.7% identity to that of Etgam22 of E. tenella. 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 E. necatrix, E. maxima and E. tenella. A specific antibody to the rEnGAM22 protein recognized the wall-forming bodies in macrogametocytes and the walls of oocysts and sporocysts. Conclusions The gene cloned from E. necatrix gametocytes is an ortholog to Etgam22 of E. tenella and presents a potential target for future recombinant subunit vaccines against coccidiosis.