Escherichia coli recombinate expression method of mycobacterium tuberculosis Rv 2837c active protein and applications thereof

A technology of Mycobacterium tuberculosis and rv2837c, which is used in the preparation of tuberculosis diagnosis and preventive preparations or medicines, recombinant expression in Escherichia coli and the field of obtaining Mycobacterium tuberculosis Rv2837c active protein, which can solve the problem of limited effect of tuberculosis antigens and no tuberculosis New vaccines and other issues

A technology of Mycobacterium tuberculosis and rv2837c, which is used in the preparation of tuberculosis diagnosis and preventive preparations or medicines, recombinant expression in Escherichia coli and the field of obtaining Mycobacterium tuberculosis Rv2837c active protein, which can solve the problem of limited effect of tuberculosis antigens and no tuberculosis New vaccines and other issues

CN105567660AActive Publication Date: 2016-05-11FOURTH MILITARY MEDICAL UNIVERSITY
3 Cites 3 Cited by

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Escherichia coli recombinate expression method of mycobacterium tuberculosis Rv 2837c active protein and applications thereof
  • Escherichia coli recombinate expression method of mycobacterium tuberculosis Rv 2837c active protein and applications thereof
  • Escherichia coli recombinate expression method of mycobacterium tuberculosis Rv 2837c active protein and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0040] Construction of Rv2837c prokaryotic expression vector:

[0041] Primers were designed according to the genome Rv2837c gene sequence of Mycobacterium tuberculosis H37Rv strain:

[0042] P1: 5'-CGGCATATGGTGACGACGATCGACCCAAG-3', NdeI site

[0043] P2: 5'-TTTAAGCTTGCCAGCTGCGCGATCTGACG-3', HindIII site

[0044] Primers were synthesized by Shanghai Sunny Biotechnology Co., Ltd. The target gene was amplified by PCR using the genomic DNA of Mycobacterium tuberculosis H37Rv strain as a template. The reaction parameters were: pre-denaturation at 95°C for 5 minutes, denaturation at 95°C for 30 seconds, renaturation at 57°C for 30 seconds, extension at 72°C for 100 seconds, 35 cycles, and 72 cycles after the last cycle. ℃ extension 5min. 1% agarose gel electrophoresis analysis showed that the 1023bp target band was amplified (see figure 1 ). The target fragment was recovered by the gel recovery kit, double-digested with NdeI and HindIII, and the digested product was recovered ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention relates to an escherichia coli recombinate expression method of mycobacterium tuberculosis Rv 2837c active protein and applications thereof. The mycobacterium tuberculosis Rv 2837c protein gene is cloned into a prokaryotic expression vector Pet28a+, and the gene is named as pET-CnpB. A Ni ion affinity chromatography method is adopted to obtain purified Rv2837c active protein. The recombinant Rv2837c protein can be used to detect the serum of animals and patients affected by mycobacterium tuberculosis. The recombinant Rv2837c protein can induce high level body fluid and cellular immunologic response of mouse, and can be used to develop novel subunit vaccine of tuberculosis. So Rv2837c protein has a good application prospect. The invention relates to the applications of the active recombinant protein in the preparation of drugs or preparations for diagnosing and preventing tuberculosis.

Description

technical field [0001] The invention belongs to the field of tuberculosis diagnostic reagents and vaccines. The invention relates to a method for recombinantly expressing and obtaining active protein of mycobacterium tuberculosis Rv2837c in Escherichia coli. The invention also relates to the application of the recombinant protein in the preparation of tuberculosis diagnosis and prevention preparations or medicines. Background technique [0002] Tuberculosis Epidemic Status [0003] Tuberculosis (TB) is a chronic infectious disease caused by Mycobacterium tuberculosis (Mtb) infection, and it is still the most serious infectious disease worldwide. According to the WHO report, about 1 / 3 of the world's population has been infected with Mycobacterium tuberculosis, and there were about 9 million new patients in 2013. Among all infectious diseases, the death rate of tuberculosis is second only to AIDS. WHO lists TB, together with AIDS and malaria, as the most important infectiou...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
11 May 2016
Publication
CN105567660A
IPC
C12N9/22; C12N15/70; G01N33/569; A61K39/04; A61P31/06
CPC
A61K39/04; A61K2039/575; C12N9/22; C12N15/70; C12N2800/101; G01N33/5695; G01N2333/922; G01N2469/20
Inventors
柏银兰; 徐志凯