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2005•Journal of Tropical MedicineRequires access

Cloning and Expression of EBA-175 of Plasmodium falciparum FCC1/HN Isolate

Xiaomin Sun

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Abstract

Objective To construct recombinant expressio n vector containing the F2domain located within regionⅡof the 175kDa Plasmodium falciparum erythrocyte binding antigen(EBA-175).Methods The 856-1848bp fragment of EBA-175was amplified with PCR.The P CR product was directionally cloned into PMD18-T.The insert was excised by BamHⅠand XhoⅠand subcloned into high expression v ector pET23a +.The recombinant protein is induced to express in BL21(DE3)transformed cells by IPTG.Results The inserted fragment in pET23a +was confirmed to be the target gene.With induction of IPTG,a new protein with the expected molecular mass of 36.4kDa was expressed.Conclusion The recombinant expression vector was c onstructed successfully,which cou ld express the recombinant EBA-175p rotein in BL21(DE3)cells.

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What this paper is about

Objective To construct recombinant expressio n vector containing the F2domain located within regionⅡof the 175kDa Plasmodium falciparum erythrocyte binding antigen(EBA-175).Methods The 856-1848bp fragment of EBA-175was amplified with PCR.The P CR product was directionally cloned into PMD18-T.The insert was excised by BamHⅠand XhoⅠand subcloned into high expression v ector pET23a +.The recombinant protein is induced to express in BL21(DE3)transformed cells by IPTG.Results The inserted fragment in pET23a +was confirmed to be the target gene.With induction of IPTG,a new protein with the expected molecular mass of 36.4kDa was expressed.Conclusion The recombinant expression vector was c onstructed successfully,which cou ld express the recombinant EBA-175p rotein in BL21(DE3)cells.

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Available abstract

Objective To construct recombinant expressio n vector containing the F2domain located within regionⅡof the 175kDa Plasmodium falciparum erythrocyte binding antigen(EBA-175).Methods The 856-1848bp fragment of EBA-175was amplified with PCR.The P CR product was directionally cloned into PMD18-T.The insert was excised by BamHⅠand XhoⅠand subcloned into high expression v ector pET23a +.The recombinant protein is induced to express in BL21(DE3)transformed cells by IPTG.Results The inserted fragment in pET23a +was confirmed to be the target gene.With induction of IPTG,a new protein with the expected molecular mass of 36.4kDa was expressed.Conclusion The recombinant expression vector was c onstructed successfully,which cou ld express the recombinant EBA-175p rotein in BL21(DE3)cells.

Key concepts: Recombinant DNA, lac operon, Molecular biology, Cloning (programming), Vector (molecular biology), Biology, Insert (composites), Molecular cloning

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Cloning and Expression of EBA-175 of Plasmodium falciparum FCC1/HN Isolate — Research Paper | ScholarLens