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Expression of Recombinant Plasmodium Falciparum Circumsporozoite Protein by Using Wheat Germ Cell-free Protein Synthesis System

Author: HanJing
Tutor: YeSiYing
School: Huazhong University of Science and Technology
Course: Pathogen Biology
Keywords: Plasmodium falciparum Circumsporozoite protein Wheat germ cell-free protein synthesis system Malaria Vaccine
CLC: R392
Type: Master's thesis
Year: 2011
Downloads: 17
Quote: 0
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Abstract


Plasmodium circumsporozoite protein (CSP) protein is evenly distributed throughout the Plasmodium sporozoite protein layer on the table is , Plasmodium falciparum sporozoite period of important protective antigens , but also malaria vaccine candidate antigen is one of the important . However, due to the complex structure of the protein , yet no suitable expression and protein synthesis system prepared as an ideal malaria vaccine recombinant CSP applications . Wheat germ cell-free protein synthesis system is a cell-free synthesis system in eukaryotic protein synthesis system is different cell protein synthesis system based on a new protein synthesis system , can be overcome so far protein synthesis in a variety of other techniques of genetic engineering shortcomings , foreign scholars have been making use of wheat germ cell-free protein synthesis system Plasmodium protein expression studies in the literature to support the Plasmodium protein expression system has unique advantages , so this issue by applying the system synthesis falciparum CSP, to investigate the feasibility of a malaria vaccine for the new malaria vaccine research and to establish a basis . Objective: wheat germ cell-free protein synthesis system having the biological activity of the expression of P. falciparum CS protein . Methods: According PlasmoDB on Richard Pf3D7 strains of P. falciparum CSP gene sequence to design a pair of primers amplified by PCR CSP gene sequences . With NcoI and SmaI DNA endonuclease digested PCR products after digestion with the same pair of pIVEX 1.3WG vector and transformed into E. coli strain DH5а obtain pIVEX WG 1.3-CSP expression plasmid . Conventional methods for the identification and plasmid DNA sequencing . The pIVEX WG 1.3-CSP plasmid was added to wheat germ cell-free protein synthesis system for expression , optimize different expression conditions , parasite proteins expressed product by partial character identification and analysis . Results : PCR successfully amplified CSP gene sequences ; successfully constructed for wheat germ cell-free system pIVEX WG 1.3-CSP eukaryotic expression plasmid ; the recombinant plasmid in wheat germ cell-free protein synthesis system successfully expressed a recombinant P. falciparum CS protein , was identified , the soluble expression product was 100% ; Conclusion: wheat germ cell-free protein synthesis system capable of expressing a completely soluble recombinant CS protein . But the expression on the cell-free system, purified recombinant protein malaria problem , still need further study.

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