Biomaterial Database

Stable transfection of Eimeria necatrix through nucleofection of second generation merozoites. 2019

Chunhui Duan, and Dandan Hu, and Xinming Tang, and Jingxia Suo, and Si Wang, and Sixin Zhang, and Geru Tao, and Chao Li, and Chaoyue Wang, and Xiaolong Gu, and Xiaoli Tang, and Guangping Huang, and Biqi Xiang, and Shaoqiang Wu, and Choukri Ben Mamoun, and Xun Suo, and Xianyong Liu

State Key Laboratory of Agrobiotechnology, China Agricultural University, Beijing, 100193, China; National Animal Protozoa Laboratory, College of Veterinary Medicine, China Agricultural University, Beijing, 100193, China.

Eimeria spp., the causative agents of coccidiosis, are the most common protozoan pathogens of chickens. Infection with these parasites can result in poor development or death of animals leading to a devastating economic impact on poultry production. The establishment of transfection protocols for genetic manipulation of Eimeria species and stable expression of genes would help advance the biology of these parasites as well as establish these organisms as novel vaccine delivery vehicles. Here, we report the selection of the first stable transgenic E. necatrix population, EnHA1, consitutively expressing the EYFP reporter following transfection of the 2nd generation merozoites with a linear DNA fragment harboring the EYFP reporter gene, the HA1 gene from the avian influenza virus H9N2 and the TgDHFR-TS selectable marker, which confers resistance to pyrimethamine. Transfected merozoites were inoculated into chickens via the cloacal route, and feces from 18 h to 72 h post inoculation were collected and subjected to subsequent serial passages, FACS sorting and pyrimethamine selection. A gradual increase in the number of EYFP-expressing sporulated oocysts was noticed with more than 90% EYFP + oocysts obtained after five passages. Immunofluorescence assay confirmed successful expression of the HA1 antigen in the EnHA1 population. The ability to genetically manipulate E. necatrix merozoites and express heterologous genes in this parasite will pave the way for possible use of this organism as a vaccine-delivery vehicle.

UI	MeSH Term	Description	Entries
D008164	Luminescent Proteins	Proteins which are involved in the phenomenon of light emission in living systems. Included are the "enzymatic" and "non-enzymatic" types of system with or without the presence of oxygen or co-factors.	Bioluminescent Protein,Bioluminescent Proteins,Luminescent Protein,Photoprotein,Photoproteins,Protein, Bioluminescent,Protein, Luminescent,Proteins, Bioluminescent,Proteins, Luminescent
D011201	Poultry Diseases	Diseases of birds which are raised as a source of meat or eggs for human consumption and are usually found in barnyards, hatcheries, etc. The concept is differentiated from BIRD DISEASES which is for diseases of birds not considered poultry and usually found in zoos, parks, and the wild.	Disease, Poultry,Diseases, Poultry,Poultry Disease
D002645	Chickens	Common name for the species Gallus gallus, the domestic fowl, in the family Phasianidae, order GALLIFORMES. It is descended from the red jungle fowl of SOUTHEAST ASIA.	Gallus gallus,Gallus domesticus,Gallus gallus domesticus,Chicken
D003048	Coccidiosis	Protozoan infection found in animals and man. It is caused by several different genera of COCCIDIA.	Besnoitiasis,Besnoitiosis,Besnoitiases,Besnoitioses,Coccidioses
D004539	Eimeria	A genus of protozoan parasites of the subclass COCCIDIA. Various species are parasitic in the epithelial cells of the liver and intestines of man and other animals.	Eimerias
D000818	Animals	Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA.	Animal,Metazoa,Animalia
D001426	Bacterial Proteins	Proteins found in any species of bacterium.	Bacterial Gene Products,Bacterial Gene Proteins,Gene Products, Bacterial,Bacterial Gene Product,Bacterial Gene Protein,Bacterial Protein,Gene Product, Bacterial,Gene Protein, Bacterial,Gene Proteins, Bacterial,Protein, Bacterial,Proteins, Bacterial
D014162	Transfection	The uptake of naked or purified DNA by CELLS, usually meaning the process as it occurs in eukaryotic cells. It is analogous to bacterial transformation (TRANSFORMATION, BACTERIAL) and both are routinely employed in GENE TRANSFER TECHNIQUES.	Transfections
D052940	Merozoites	Uninuclear cells or a stage in the life cycle of sporozoan protozoa. Merozoites, released from ruptured multinucleate SCHIZONTS, enter the blood stream and infect the ERYTHROCYTES.	Merozoite
D017930	Genes, Reporter	Genes whose expression is easily detectable and therefore used to study promoter activity at many positions in a target genome. In recombinant DNA technology, these genes may be attached to a promoter region of interest.	Reporter Genes,Gene, Reporter,Reporter Gene