Enhanced Secretion of Biologically Active Murine Interleukin-12 by Lactococcus lactis
APPLIED AND ENVIRONMENTAL MICROBIOLOGY
Authors: Fernandez, Antonio; Horn, Nikki; Wegmann, Udo; Nicoletti, Claudio; Gasson, Michael J.; Narbad, Arjan
Abstract
The novel signal peptide SLPmod was used for the secretion of murine interleukin-12 (mIL-12) by Lactococcus lactis. A >4-fold increase in secretion was observed when SLPmod was used instead of the Usp45-derived secretion signal. Oral delivery of this cytokine using the autoinducible host L. lactis FI5876 utilizing SLPmod resulted in a significant increase in mIL-12 plasma levels in mice.
The production of SPusp45-MSP-1(19) gene construct and its recombinant protein in Lactococcus lactis to be used as a malaria vaccine
MEDICAL JOURNAL OF INDONESIA
Authors: Kusuma, Amino V. A.; Mustopa, Apon Z.; Mustafawi, Wike Z.; Suharsono
Abstract
Background: Merozoite surface protein 1 (MSP-1) is a major protein used by the Plasmodium during red blood cells invasion in malaria. MSP-1(19), 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-1(19) gene construct and the recombinant protein in Lactococcus lactis. Methods: Usp45-MSP-1(19), derived from codon optimization and the synthetic gene, was inserted into the pMAT cloning vector. A vector expressing MSP-1(19) included usp45 has been constructed by the manipulation of recombinant DNA using restriction enzymes. The MSP-1(19) 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-1(19) 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-1(19) 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-1(19) gene was successfully inserted into the multiple cloning sites of the pNZ8148 expression vector and the MSP-1(19) protein expressed in the NICE system of the L. lactis host cell.
COA
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