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Gonococcus immunodepression mutant gene delta opaI and mutation method thereof

A technology of immunosuppression and mutating genes, which is applied in the field of microbiology to achieve the effect of eliminating immunosuppression, solving bottleneck problems, and preventing re-infection

Inactive Publication Date: 2012-07-11
CHANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The present invention provides a mutation opa I method and mutation gene, thereby eliminating the immunosuppressive effect of Neisseria gonorrhoeae, after the mutant strain enters the body, it can stimulate a normal immune response, which solves the bottleneck problem in the development of Neisseria gonorrhoeae whole-cell vaccine

Method used

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  • Gonococcus immunodepression mutant gene delta opaI and mutation method thereof
  • Gonococcus immunodepression mutant gene delta opaI and mutation method thereof
  • Gonococcus immunodepression mutant gene delta opaI and mutation method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0024] (one) opa Cloning of I gene and amplification of flanking regions on both sides

[0025] 1, opa Cloning of the I gene

[0026] Neisseria gonorrhoeae genomic DNA as template, F: 5'CG GGATCC GCGGGTGAAGGCAATGGC3'( Bam H I) (SEQ ID NO.1) and R: 5'CCC GGATCC GAAGCGGTAGCGCACGCC3' ( Bam The sequence of H I) (SEQ ID NO.2) is a primer amplified by PCR in the Neisseria gonorrhoeae genome opa I gene ( figure 1 ), the amplified product was sequenced, and the sequence is shown in SEQ ID NO.3. In SEQ ID NO.3, the two ends are Bam H I recognition site.

[0027] 2. opa Amplification of flanking regions upstream and downstream of I

[0028] the above opa The I gene amplification product is connected with the T vector pMD-19T, and the connection product is used as a template to amplify the containing opa A linear fragment of the flanking regions on both sides of the I gene, the agarose gel electrophoresis analysis of the linear fragment is as follows figure 2 , the ...

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Abstract

The invention relates to the field of microorganism, in particular to a gonococcus immunodepression mutant gene delta opaI and a mutation method thereof. A nucleotide sequence of the gonococcus immunodepression mutant gene delta opaI is described as SEQ ID NO.10. The mutant gene is obtained through the following method: gonococcus genome deoxyribonucleic acid (DNA) is used as a template, flank area DNA fragments on the upstream and the downstream of the opaI gene are amplified through a polymerase chain reaction (PCR), a kanamycin resistant gene Kanr is inserted between the gonococcus genome DNA and the opaI gene, and the mutant gene delta opaI is obtained.

Description

technical field [0001] The invention relates to the field of microbiology, in particular to a gonorrhea immunosuppressive mutation gene and a mutation method. Background technique [0002] The natural host of Neisseria gonorrhoeae is human beings, which lead to the sexually transmitted disease gonorrhea after infection, which can cause a series of lesions, including local infections (such as urethritis, epididymitis, cervicitis, salpingitis, etc.) and systemic disseminated infections. Lesions in the fallopian tubes and pelvic cavity can lead to serious sequelae such as infertility or ectopic pregnancy. Gonorrhea patients also greatly increased the chance of AIDS. The incidence of gonorrhea is high, with approximately 62 million new cases worldwide each year. The increasingly serious drug resistance of gonorrhea has brought more and more difficulties to the treatment of gonorrhea. Therefore, developing an effective vaccine is one of the key directions in the prevention and...

Claims

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Application Information

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IPC IPC(8): C12N15/31C12N15/09
Inventor 王小兵
Owner CHANGZHOU UNIV
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