Molecular cloning, sequencing, and expression of Eimeria tenella HSP70 partial gene

Abstract

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.

Eimeria falciformis secretes extracellular vesicles to modulate proinflammatory response during interaction with mouse intestinal epithelial cells

“…2 ) suggests that specific protein could be secreted via two or more secretory pathways during host-parasite interactions. The secretion of Hsp70 by Eimeria species is associated with sporozoite formation, pathogenicity, protective immunity and response to stress [ 5 , 58 , 59 ]. Hsp70 has also been reported among EV proteins from C. parvum, N. caninum , T. gondii , Leishmania and Trypanosoma species [ 7 ].…”

Eimeria proteins: order amidst disorder

“…However, heterogeneity or nucleotide diversity of protein genes could vary in different isolates [ 60 ]. Other prominent protein genes that have been mapped are hsp90 [ 57 ], hsp70 [ 61 ] as well as protease genes in which > 40 are already identified in E. tenella genome [ 62 ]. Although eimerian developmental stages share many transcriptional and translational products [ 63 ], each stage possibly has varying threshold of gene expression [ 64 ] and translational profiles [ 55 ].…”

“…Despite this, Hsp70 plays an important role in the formation of sporocysts and sporozoites [ 61 ] (Table 1 ). A dose-dependent inhibition of Hsp70 by quercetin inhibited the formation of syneptonema complex and haploidy in E. tenella sporozoite suggesting that Hsp70 could act as sentinel for assembly and disassembly of other proteins during developmental transition [ 141 ].…”