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Chemical Synthesis, Expression and Application of the Gene Fragment of Streptococcus Pneumoniae Surface Adhesin-a

A Streptococcus pneumoniae, chemical synthesis technology, applied in the field of genetic engineering technology and detection reagents, can solve problems such as being unsuitable for rapid detection and long experimental time, and achieve the effect of avoiding interference and easy purification

Active Publication Date: 2017-02-15
中国人民解放军东部战区疾病预防控制中心
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  • Application Information

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Problems solved by technology

However, these methods more or less have shortcomings such as long experiment time and not suitable for on-site rapid detection.

Method used

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  • Chemical Synthesis, Expression and Application of the Gene Fragment of Streptococcus Pneumoniae Surface Adhesin-a
  • Chemical Synthesis, Expression and Application of the Gene Fragment of Streptococcus Pneumoniae Surface Adhesin-a
  • Chemical Synthesis, Expression and Application of the Gene Fragment of Streptococcus Pneumoniae Surface Adhesin-a

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Embodiment Construction

[0047] Detailed description of the embodiment of the present invention:

[0048] Analysis, Gene Synthesis and Expression of Adhesin A Antigen Epitope of Streptococcus pneumoniae

[0049] By computer analysis of the entire amino acid sequence of the PsaA protein of Streptococcus pneumoniae, the strong antigenic epitope in the PsaA protein is screened out, and the codons preferred by bacteria are selected to chemically synthesize a new gene fragment of the strong antigenic epitope of the PsaA protein. Cloning the gene fragment into the plasmid pGEX4T-2 Bam HI / xho The I site, consistent with the translation frame of the initiation 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 PsaA protein were obtained by screening, and the expressed PsaA protein accounted for about 30% of the total bacterial protein.

[0050] Materials and Methods

[0051] 1. Str...

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Abstract

The invention provides a gene segment of chemosynthetic streptococcus pneumoniae surface adhesin A as well as expression and application of the gene segment, and relates to the fields of a genetic engineering technology and diagnostic reagents. A bran-new gene sequence of an epitope is chemically synthesized by screening out a strong antigen epitope and total 197 amino acids from the 84th amino acid to the 280th amino acid in streptococcus pneumoniae adhesin A through computer analysis and selecting a codon which a procaryotic organism prefers. The genetic engineering technology is used for expressing the gene segment and preparing a strong antigen epitope segment of the streptococcus pneumoniae adhesin A. Expressed protein can be used for vaccine research and development, detection of streptococcus pneumoniae infection antibodies and preparation of monoclonal antibodies and polyclonal antibodies.

Description

technical field [0001] The present invention chemically synthesizes the gene fragment of Streptococcus pneumoniae surface adhesin A and its expression and application relate to a new gene fragment of chemically synthesizing Streptococcus pneumoniae surface adhesin A (PsaA protein), which is prepared by using genetic engineering technology Recombinant protein. Through computer analysis, the fragments of Streptococcus pneumoniae PsaA protein containing strong epitopes are screened out, the codons preferred by prokaryotes are selected, and a new gene sequence is chemically synthesized, expressed by genetic engineering technology, and 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. Background technique [0002] Streptococcus pneumoniae (Streptococcus pneumoniae) referred to as pneum...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/31C12N15/70C07K14/315C07K16/12G01N33/569C12R1/465
Inventor 李越希蔡冉徐悦玥许桂丽周洁李素芹潘英李丙军陈乐如张素芬马颖袁敬宇
Owner 中国人民解放军东部战区疾病预防控制中心
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