The production of SPusp45-MSP-119 gene construct and its recombinant protein in Lactococcus lactis to be used as a malaria vaccine

  • Amino V.A. Kusuma Research Center for Biotechnology, Indonesian Institute of Science (LIPI), Bogor
  • Apon Z. Mustopa Research Center for Biotechnology, Indonesian Institute of Science (LIPI), Bogor
  • Wike Z. Mustafawi Research Center for Biotechnology, Indonesian Institute of Science (LIPI), Bogor
  • Suharsono Suharsono Research Center for Bioresources and Biotechnology, Bogor Agricultural University, Bogor
DOI: https://doi.org/10.13181/mji.v26i4.2162
Keywords: Lactococcus lactis, malaria, merozoite surface protein 1, nisin, usp45-MSP-119
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Abstract

Background: Merozoite surface protein 1 (MSP-1) is a major protein used by the Plasmodium during red blood cells invasion in malaria. MSP-119, one of MSP-1 is highly conserved, and it is a potential malaria vaccine candidate because the monoclonal antibodies are capable blocking erythrocyte invasion in vitro. The aim of this study was to produce MSP-119 gene construct and the recombinant protein in Lactococcus lactis.

Methods: Usp45-MSP-119, derived from codon optimization and the synthetic gene, was inserted into the pMAT cloning vector. A vector expressing MSP-119 included usp45 has been constructed by the manipulation of recombinant DNA using restriction enzymes. The MSP-119 protein was expressed to 45% ammonium sulfate precipitation and purified using Sephadex-G50 gel filtration chromatography. The expressed protein was characterized by SDS-PAGE and dot blot.

Results: usp45-MSP-119 gene was amplified using specific primers and inserted into the multiple cloning sites in the expression vector pNZ8148 with size 3,538 bp as a recombinant vector. The protein of MSP-119 was successfully expressed in L. lactis with molecular weight of 10.45 kDa. The dot blot was tested in 3 different comparisons between the host cells, non-induced cells, and induced cells with 10 ng/ml nisin. The results showed that 10 ng/ml nisin gave a positive reaction as detected by dot blot assay.

Conclusion: This study confirmed that the usp45-MSP-119 gene was successfully inserted into the multiple cloning sites of the pNZ8148 expression vector and the MSP-119 protein expressed in the NICE system of the L. lactis host cell.

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Published
2018-02-14
How to Cite
1.
Kusuma AV, Mustopa AZ, Mustafawi WZ, Suharsono S. The production of SPusp45-MSP-1<sub>19</sub> gene construct and its recombinant protein in <em>Lactococcus lactis</em&gt; to be used as a malaria vaccine. Med J Indones [Internet]. 2018Feb.14 [cited 2024May2];26(4):261-9. Available from: http://mji.ui.ac.id/journal/index.php/mji/article/view/2162
Issue
Vol. 26 No. 4 (2017): December
Section
Basic Medical Research

Copyright (c) 2018 Amino V.A. Kusuma, Apon Z. Mustopa, Wike Z. Mustafawi, Suharsono Suharsono

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