The production of SPusp45-MSP-119 gene construct and its recombinant protein in Lactococcus lactis to be used as a malaria vaccine
Abstract: 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.
Keywords: Lactococcus lactis;
malaria; merozoite surface protein 1; nisin; usp45-MSP-119
Author: Amino V.A. Kusuma,
Apon Z. Mustopa, Wike Z. Mustafawi, Suharsono
Journal Code: jpkedokterangg170236
