title: Laboratory Scale Production of Recombinant Haa86 Tick Protein in Pichia pastoris and in Escherichia coli System.
abstract: The commercial recombinant Bm86-based vaccines against Rhipicephalus (Boophilus) microplus in Australia (TickGARD™, TickGARD plus™) and in Cuba (Gavac™) provided significant impetus to researchers globally to work on anti-tick vaccines. The Bm86 homologue of Hyalomma anatolicum anatolicum Izatnagar isolate, rHaa86, is considered a potent vaccine candidate against Hyalomma tick species, transmitting animal and human diseases. The two expression systems, prokaryotic, using bacterial expression plasmid vectors and Escherichia coli, and eukaryotic, using methylotrophic yeast Pichia pastoris and yeast expression plasmid vectors, are used in the laboratory level production of rHaa86. Unlike proteins expressed in prokaryotic system, eukaryotic system-expressed proteins are glycosylated and may be a requisite for proper immunogenicity. Here, the protocol for laboratory scale expression of rHaa86 antigen in E. coli and P. pastoris for development of an anti-Hyalomma tick vaccine is described.
authors.affiliation: Entomology Laboratory, Parasitology Division, Indian Veterinary Research Institute, Izatnagar, Bareilly, Uttar Pradesh, 243122, India.|Department of Veterinary Parasitology, College of Veterinary Science and Animal Husbandry, Junagadh Agricultural University, Junagadh, 362001, Gujarat, India.
authors.forename: Binod
authors.initials: B
authors.lastname: Kumar
authors.affiliation: Entomology Laboratory, Parasitology Division, Indian Veterinary Research Institute, Izatnagar, Bareilly, Uttar Pradesh, 243122, India.|Department of Veterinary Parasitology, Madras Veterinary College, Chennai, Tamil Nadu, India.
authors.forename: Azhahianambi
authors.initials: A
authors.lastname: P
authors.affiliation: Entomology Laboratory, Parasitology Division, Indian Veterinary Research Institute, Izatnagar, Bareilly, Uttar Pradesh, 243122, India. sghoshtick@gmail.com.
authors.forename: Srikant
authors.initials: S
authors.lastname: Ghosh
mesh_terms: D000818:Animals; D060829:Arthropod Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics*; D005818:Genetic Engineering / Q000379:methods*; D026863:Ixodidae; D007753:Laboratories; D010843:Pichia / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D017422:Sequence Analysis, DNA
chemical_list: D060829:Arthropod Proteins; D018076:DNA, Complementary; D011994:Recombinant Proteins
keywords: Anti-Hyalomma vaccine; Eukaryotic expression; Laboratory scale production; Prokaryotic expression; rHaa86