title: Molecular cloning, sequencing, and expression of Eimeria tenella HSP70 partial gene.
abstract: Members of the Eimeria genus are protozoan parasites of the subphylum Apicomplexa (Eimeriidae family), and belong to the coccidia group. Eimeria tenella is one of the most pathogenic species owing to its ability to penetrate the mucosa, and cause inflammation and damage. It is an obligate intracellular parasite that causes disease by destroying the host cells during multiplication. Heat shock protein 70 (HSP70) is a molecular chaperone that prevents cellular stress. The objective of this study was to clone, sequence, and express E. tenella HSP70 protein. After selecting the region of highest hydrophilicity in the hsp70 gene, we cloned complementary DNA (cDNA) into a pTrcHis2-TOPO vector and transformed it into TOP10 Escherichia coli cells; after induction, the bacteria expressed a 23-kDa protein with insoluble expression levels of approximately 5 mg/L. In summary, the partial hsp70 gene was successfully expressed in E. coli, producing a 23-kDa protein under insoluble conditions, and the antigen characteristics predicted by hydrophilicity analysis suggest the development of a vaccine for use in avian coccidiosis.
authors.affiliation: Laboratório de Protozoologia, Departamento de Medicina Veterinária Preventiva, Universidade Estadual de Londrina, Londrina, PR, Brasil.
authors.forename: A L G
authors.initials: AL
authors.lastname: Bogado
authors.affiliation: Laboratório de Protozoologia, Departamento de Medicina Veterinária Preventiva, Universidade Estadual de Londrina, Londrina, PR, Brasil.
authors.forename: G F
authors.initials: GF
authors.lastname: Martins
authors.affiliation: Laboratório de Protozoologia, Departamento de Medicina Veterinária Preventiva, Universidade Estadual de Londrina, Londrina, PR, Brasil.
authors.forename: J P
authors.initials: JP
authors.lastname: Sasse
authors.affiliation: Laboratório de Protozoologia, Departamento de Medicina Veterinária Preventiva, Universidade Estadual de Londrina, Londrina, PR, Brasil.
authors.forename: J da S
authors.initials: JD
authors.lastname: Guimarães
authors.affiliation: Laboratório de Protozoologia, Departamento de Medicina Veterinária Preventiva, Universidade Estadual de Londrina, Londrina, PR, Brasil joaoluisgarcia10@gmail.com.
authors.forename: J L
authors.initials: JL
authors.lastname: Garcia
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002645:Chickens; D016786:Eimeria tenella / Q000235:genetics*; D004926:Escherichia coli; D015870:Gene Expression; D018840:HSP70 Heat-Shock Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D057927:Hydrophobic and Hydrophilic Interactions; D015800:Protozoan Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D017422:Sequence Analysis, DNA
chemical_list: D018840:HSP70 Heat-Shock Proteins; D015800:Protozoan Proteins