MSP-1 protein preparations from plasmodium
A MSP-1, protein technology, applied in medical preparations containing active ingredients, introduction of foreign genetic material using vectors, resistance to vector-borne diseases, etc., can solve problems such as low production efficiency of p83/30 subunits
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Embodiment 1
[0170] Example 1: Mutation of Nucleic Acid Sequence Encoding MSP-1 Protein Subunit
[0171] The ribosome binding site in prokaryotes is also called the "Shine-Dalgarno sequence" and consists of a purine-rich sequence about 10 nucleotides upstream of the start codon. The ribosome binding site sequence for expression of the recombinant MSP-1 protein in plasmid pZE23 / d-83 / 30 corresponds to the "Shine-Dalgano" consensus sequence (Shine & Dalgarno (1975) Nature 254, 34-8) and has been previously described. Its ability to drive gene expression in E. coli is described (Lutz & Bujard (1997) Nucleic Acids Res. 25, 1203-10). Due to the fact that translation of mRNA into protein in prokaryotes occurs while transcription is still in progress, the secondary structure of mRNA may affect the recognition of mRNA by ribosomes and thus the rate of protein synthesis.
[0172] According to the above, the analysis of the mRNA structure in silico simulations was carried out through the RNA struc...
Embodiment 2
[0176] Example 2: Comparison of non-mutated and mutated MSP-1 subunit expression
[0177] will contain pZE23 / d-83 / 30 (corresponding to the unmodified sequence of the p83 / 30 fragment of SEQ ID NO: 6 or 7, wherein the ribosome binding sites are equivalent) or pZE23 / d-mut83 / 30 (corresponding to Escherichia coli W3110-Z2 (modified sequence of p83 / 30 fragment of SEQ ID NO: 8 or 9, wherein the ribosome binding site has been equivalently modified) was grown in Superbroth medium at 37° C. to a cell density of OD500 = 0.5. MSP-1 expression was induced by adding IPTG (1 mM) and the bacteria were grown for an additional 4 hours. Aliquots of total bacterial extracts were taken at induction (Oh) and 2 (2h) and 4 hours (4h) thereafter. Total cell extracts were analyzed by SDS-PAGE and Coomassie staining. The results obtained are shown in figure 2 .
Embodiment 3
[0178] Example 3: Production of heterodimeric MSP-1 from p83 / 30 and p38 / 42
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