title: Cloning, overexpression and characterization of soluble 42kDa fragment of merozoite surface protein-1 of Plasmodium vivax.
abstract: Plasmodium vivax represents the second most prevalent malaria species of major public health importance and the global eradication of malaria requires the development of vaccines to prevent infection. The lack of in vitro culture and a suitable animal model for P. vivax malaria are the major problems for the delay in developing a functional vivax vaccine. A number of antigens have been identified for P. vivax as potential malaria vaccine candidates and among these 42kDa fragment of merozoite surface protein-1 (MSP-142) is one of most promising antigen of asexual blood stage. In most of the earlier studies, the MSP-142 of malaria parasites was expressed as insoluble protein in inclusion bodies and it is difficult to get purified protein in conformation form. In the present study, we have cloned, overexpressed and characterized the 42kDa fragment of P. vivax MSP-1 as soluble protein in Escherichiacoli. The 42kDa gene fragment of P. vivax MSP-1 was PCR amplified using specific primers, sequenced and subcloned into pTriEx-4 expression vector. The optimum expression of recombinant P. vivax protein was obtained in SOC growth medium by inducing with 0.2mM IPTG at 37°C for 4h. The SDS-PAGE analysis showed a fusion protein of 55kDa and about 80% was present in soluble form. The purified P. vivax MSP-142 was characterized and found to be correctly folded and in conformation form as evident by CD spectroscopy, presence of 1 free -SH group and the reactivity with reduction sensitive conformational monoclonals against P. vivax MSP-142.
authors.affiliation: Division of Parasitology, CSIR-Central Drug Research Institute, Lucknow 226031, India; Department of Biochemistry, Lucknow University, Lucknow, India.
authors.forename: Inayat Hussain
authors.initials: IH
authors.lastname: Sheikh
authors.affiliation: Amity University Uttar Pradesh, Lucknow Campus, Lucknow 226010, India.
authors.forename: Deep C
authors.initials: DC
authors.lastname: Kaushal
authors.affiliation: Division of Parasitology, CSIR-Central Drug Research Institute, Lucknow 226031, India.
authors.forename: Vandana
authors.initials: V
authors.lastname: Singh
authors.affiliation: Division of Parasitology, CSIR-Central Drug Research Institute, Lucknow 226031, India.
authors.forename: Niraj
authors.initials: N
authors.lastname: Kumar
authors.affiliation: Department of Biochemistry, Lucknow University, Lucknow, India.
authors.forename: Deepak
authors.initials: D
authors.lastname: Chandra
authors.affiliation: Division of Parasitology, CSIR-Central Drug Research Institute, Lucknow 226031, India. Electronic address: nuzhatkaushal@hotmail.com.
authors.forename: Nuzhat A
authors.initials: NA
authors.lastname: Kaushal
mesh_terms: D000818:Animals; D001483:Base Sequence; D006801:Humans; D017780:Malaria Vaccines / Q000235:genetics; D016780:Malaria, Vivax / Q000235:genetics / Q000276:immunology / Q000517:prevention & control*; D020066:Merozoite Surface Protein 1 / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D010966:Plasmodium vivax / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology
chemical_list: D017780:Malaria Vaccines; D020066:Merozoite Surface Protein 1; D011994:Recombinant Proteins
keywords: Malaria; Malaria vaccine; Merozoite surface protein-1; Plasmodium vivax