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Recombination protein for preventing human trachoma bedsnia infestation and its use

A Chlamydia trachomatis and recombinant protein technology, applied in the field of genetic engineering, can solve problems such as unrealistic immune protection

Inactive Publication Date: 2006-03-01
BEIJING HYDVAX BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For now, it is unrealistic to use this method to achieve immune protection, but it suggests that immunity to non-viable bacterial infection can also achieve immune protection

Method used

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  • Recombination protein for preventing human trachoma bedsnia infestation and its use
  • Recombination protein for preventing human trachoma bedsnia infestation and its use
  • Recombination protein for preventing human trachoma bedsnia infestation and its use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0073] Example 1 Artificial construction of epitope gene

[0074] Connect the selected epitope genes on the main outer membrane protein together, and construct an EcoRI restriction site at the amino-terminus, and a HindIII restriction site at the carboxy-terminus, to obtain Chlamydia trachomatis that can activate human T cells A gene encoding a major outer membrane protein epitope (hereinafter referred to as "chlamy").

[0075] The specific method is as follows: artificially construct the selected epitope gene through 3 rounds of PCR cycles. Primer 3 and primer 4 serve as templates and primers for the first round of PCR; then use the first round of PCR products as templates and primers 2 and 5 as primers for the second round of PCR; then use the second round of PCR products as As a template, primers 1 and 6 were used as primers for the third round of PCR.

[0076] The primer sequences are as follows:

[0077] primer1 (chlamy-1)

[0078] 5'GAATTCCCGGCATACGGTCGTCATATGCAGGATG...

Embodiment 2

[0124] Example 2 TA Cloning of Epitope Genes

[0125] The PCR product was cloned by TA cloning method (J. Sambrook, Cold Spring Harbor Laboratory Press, Molecular cloning, 1989), and the vector of TA cloning was PMD 18-T Vector (TakaRa Company). The specific method is as follows:

[0126] 1 DNA recovery:

[0127] 1. Electrophoresis of the third-round PCR product with 2% agarose gel (1×TAE, 150-200mA, 0.5 hour);

[0128] 2. Cut the gel containing the DNA fragment from the agarose gel and put it into a centrifuge tube;

[0129] 3. Add 3 times the volume of sol solution (Beijing Dingguo Company), and bathe in water at 45-55°C for 5-10 minutes to completely melt the glue;

[0130] 4. Add 10ul glass milk (Beijing Dingguo Company), flick the bottom of the tube to mix well, and then put it in a water bath at 45-55℃ for 5-10min, during which time mix every 2-3min;

[0131] 5. Centrifuge at 5000g for 60sec, discard the supernatant;

[0132] 6. Add 400ul rinse solution, flick the b...

Embodiment 3

[0149] Obtain the coding gene of Mycobacterium tuberculosis 65KD heat shock protein (BCG HSP65) and clone it into pET28(+) expression vector

[0150] BCG Mycobacterium tuberculosis was obtained from Changchun Institute of Biological Products. Use Sutong potato medium (Starch Potato Code NO C250-1 Beijing Dingguo Biology) to cultivate BCG Mycobacterium tuberculosis at a temperature of 37-39°C, and the grown BCG Mycobacterium tuberculosis will appear as dry wrinkled light yellow bacteria membrane. The pellicles were collected, from which BCG M. tuberculosis genomic DNA was extracted.

[0151] The method of extracting Mycobacterium tuberculosis genomic DNA refers to Molecular Cloning book (J.Sambrook, separate high-molecular-weight DNA (Isolation of high-molecular-weight DNA from mammalian cells) from mammals), 9.16-9.22, Cold Spring Harbor Laboratory Press, Molecular cloning, 1989).

[0152] Isolation of heat shock protein 65 (HSP65) structural gene from Mycobacterium tubercu...

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Abstract

A recombinant protein vaccine for preventing the infection of human chlamydia trachomatis, which is a fusion protein recombined by linking the heat shock protein 65 of BCG vaccine with the epitope of main outer membrane protein of chlamydia trachomatis, the nucleotide sequence for coding it, the expression carrier containing said nucleotide sequence, the host cell containing said expression carrier, and the process for preparing said recombinant protein vaccine are disclosed.

Description

technical field [0001] The present invention relates to a field of genetic engineering, in particular to a genetic engineering recombinant protein vaccine (hereinafter sometimes referred to as "genetic engineering recombinant protein"), in particular to a vaccine for the prevention of human Chlamydia trachomatis infection, especially the prevention of genitourinary system infection Recombinant protein vaccine; the nucleotide sequence encoding this recombinant protein vaccine (hereinafter sometimes referred to as "gene"); the expression vector containing the nucleotide sequence; the host cell containing the expression vector, and the recombinant protein vaccine Preparation method; the present invention also relates to the use of the genetically engineered recombinant protein in the preparation of vaccine products for preventing human Chlamydia trachomatis infection and vaccine products containing the genetically engineered recombinant protein. Background technique [0002] Ch...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/118A61K38/16C12N15/09C12N15/31C12N15/62C12N15/79C12N15/63A61P13/00A61P31/00A61K39/00C07K14/295
CPCC07K14/295A61K39/00C07K2319/00A61P13/00A61P31/00
Inventor 王丽颖杨思睿于永利
Owner BEIJING HYDVAX BIOTECH
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