Chemosynthetic extracellular region gene fragment of streptococcus pneumonia PspA protein, and expression and application thereof

A Streptococcus pneumoniae and gene fragment technology, applied in the field of genetic engineering technology, can solve problems such as large variability in the alpha helix region, and achieve the effects of avoiding interference and easy purification

Active Publication Date: 2014-06-04
李越希
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  • Abstract
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Problems solved by technology

Studies have shown that PspA protein has extensive homology in strains of different serotypes, but there is a large variability in its α-helical region

Method used

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  • Chemosynthetic extracellular region gene fragment of streptococcus pneumonia PspA protein, and expression and application thereof
  • Chemosynthetic extracellular region gene fragment of streptococcus pneumonia PspA protein, and expression and application thereof
  • Chemosynthetic extracellular region gene fragment of streptococcus pneumonia PspA protein, and expression and application thereof

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Embodiment Construction

[0048] Detailed description of the embodiment of the present invention:

[0049] Epitope Analysis, Gene Synthesis and Expression of Surface Protein A Antigen of Streptococcus pneumoniae

[0050] Through the computer analysis of the entire amino acid sequence of the PspA protein of Streptococcus pneumoniae, the strong antigenic epitope in the PspA protein is screened out, and the codons preferred by the bacteria are selected to chemically synthesize a new gene fragment of the strong antigenic epitope of the PspA protein. Cloning the gene fragment into the plasmid pGEX4T-2 Bam HI / Eco The RI site, consistent with the translation frame of the start codon on the vector, can express a fusion protein. The recombinant plasmid was transformed into Escherichia coli BL21 (DE3), and the engineered bacteria that highly expressed PspA protein were obtained by screening, and the expressed PspA protein accounted for about 30% of the total bacterial protein.

[0051] Materials and Me...

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Abstract

The invention relates to the fields of genetic engineering technology and diagnostic reagents and provides a chemosynthetic extracellular region gene fragment of streptococcus pneumonia PspA protein and expression and application thereof. According to the invention, strong epitope in streptococcus pneumonia PspA protein is screened through computer analysis, the fragment consisting of the 33rd amino acid to the 109th amino acid, altogether 77 amino acids, is used, codon preferred by prokaryotes is selected, and a brand new gene sequence of the epitope is chemically synthesized; and through usage of genetic engineering technology, the gene fragment is expressed, and the fragment of the strong epitope in streptococcus pneumonia PspA protein is prepared. The expressed protein can be used for research and development of vaccines, detection of streptococcus pneumonia infection antibodies and preparation of monoclonal antibodies and polyclonal antibodies.

Description

technical field [0001] The chemically synthesized Streptococcus pneumoniae PspA protein extracellular region gene fragment and its expression and application relate to the expression, purification and application of a Streptococcus pneumoniae surface protein A antigen fragment, through chemical synthesis of Streptococcus pneumoniae surface protein A ( PspA protein) brand-new gene fragment, using genetic engineering technology to prepare recombinant protein. Through computer analysis, the fragments of Streptococcus pneumoniae PspA protein containing strong epitopes are screened out, the codons preferred by prokaryotes are selected, and a new gene sequence is chemically synthesized and expressed by genetic engineering technology. The expressed protein can be used for Streptococcus pneumoniae infection antibodies The detection and preparation of monoclonal antibodies, etc., the present invention relates to the field of genetic engineering technology and detection reagents. Back...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/31C12N15/70C07K14/315C07K1/22C07K16/12G01N33/68C12R1/46
Inventor 李越希陈乐如蔡冉周洁张素芬许桂丽马颖袁敬宇李素芹潘英李丙军徐悦玥
Owner 李越希
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