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

Inactive Publication Date: 2011-08-03
H・布加德
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0018] An additional problem encountered when constructing heterodimers of p83 / 30 and p38 / 42 in a manner strictly analogous to Kauth et al. is the very inefficient production of p83 / 30 subunits in E. coli

Method used

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  • MSP-1 protein preparations from plasmodium
  • MSP-1 protein preparations from plasmodium
  • MSP-1 protein preparations from plasmodium

Examples

Experimental program
Comparison scheme
Effect test

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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PUM

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Abstract

The invention relates to a merozoite surface protein 1 (''MSP-1'') preparation of Plasmodium falciparum, said MSP-1 preparation comprising a) a purified fragment p83 / 30 of the gp190 / MSP-1 from Plasmodium without heterologous sequences, and b) a purified fragment p38 / 42 of the gp190 / MSP-1 from Plasmodium without heterologous sequences.

Description

technical field [0001] The present invention relates to merozoite surface protein 1 (MSP-1) from the genus Plasmodium, a method for the preparation of a modified nucleic acid encoding its p83 / 30 protein subunit, a corresponding modified nucleic acid, a vector comprising it, a corresponding MSP -1 p83 / 30 subunit, method for preparing heterodimeric MSP-1, corresponding heterodimeric MSP-1 for use as a vaccine, these heterodimeric MSP-1 proteins are used Use in the preparation of medicaments for the prevention and / or treatment of malaria and vaccines comprising any of the aforementioned nucleic acid, vector or protein products. Background technique [0002] Malaria is one of the most devastating infectious diseases in the world. According to estimates from the World Health Organization (WHO), 400 to 900 million disease occurrences are recorded each year. According to information from the Multilateral Initiative against Malaria (MIM), between 700,000 and 2.7 million people die...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/455C12N15/67A61K39/015
CPCA61K2039/55572A61K2039/55555A61K39/015A61K2039/55561A61K2039/55505A61K2039/55516A61K2039/55566C07K14/445A61P33/02Y02A50/30
Inventor H·布加德R·鲁兹C·易普U·沃尔比C·考特
Owner H・布加德
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