Mycobacterium tuberculosis 38KD protein DNA extraction and recombinant vector construction expressing method

A technology of Mycobacterium tuberculosis and recombinant vectors, which is applied in the fields of protein prokaryotic function expression, recombinant vector construction, and DNA extraction of pathogenic bacteria proteins, and can solve problems such as 38KD protein research reports that have not been seen.

Inactive Publication Date: 2018-11-06
AFFILIATED HOSPITAL OF ZUNYI MEDICAL COLLEGE
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Problems solved by technology

Due to the complexity of the components of Mycobacterium tuberculosis, there are many protein antigens, and each protein has different effects on dendritic cells. As one of the important protein components, 38KD protein can cause the body's immune response! However, there are no relevant research reports on 38KD protein and dendritic cells; what impact will it have on dendritic cells, let’s learn about 38KD protein

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  • Mycobacterium tuberculosis 38KD protein DNA extraction and recombinant vector construction expressing method
  • Mycobacterium tuberculosis 38KD protein DNA extraction and recombinant vector construction expressing method
  • Mycobacterium tuberculosis 38KD protein DNA extraction and recombinant vector construction expressing method

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Embodiment Construction

[0027] Mycobacterium tuberculosis 38KD protein DNA extraction, recombinant vector construction, protein expression method of the present invention are described as follows again:

[0028] Steps and methods

[0029] 1. Amplification and identification of 38KD protein cDNA

[0030] MTB Genomic DNA Extraction: Use alkaline lysis method to extract H37Rv genomic DNA. For details, please refer to the instructions of Biyuntian DNA Extraction Kit. Using the extracted MTB-DNA as a template, MTB-38KD Primer1 and Primer2 were used as primers (the 5' ends were introduced with restriction enzymes BamH I and Hind Restriction site), 38KD upstream guide 5'-CGCGGATCCATTCGTTTGCATACGCT, (underlined part is BamH I restriction site) Primer2: 38KD downstream guide 5'-CCCAAGCTTCTAGCTGGAAATCGTCGC, (underlined is Hind Restriction sites) were purified by PAG, and the OD value of each tube was 1), and the target fragment was amplified by PCR using the extracted target gene as a template. Amplificat...

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Abstract

A mycobacterium tuberculosis 38KD protein DNA extraction and recombinant vector construction expressing method comprises the steps of amplification and identification of 38KD protein cDNA, construction of a prokaryotic expression vector MTB38KD, expression and identification of a MTB38KD protein and purification of an 8KD protein.

Description

technical field [0001] The invention relates to prokaryotic function expression methods such as Mycobacterium tuberculosis 38KD protein DNA extraction, recombinant vector construction, and protein expression, and corresponding equipment. It belongs to the technical field of pathogenic bacteria protein DNA extraction, recombinant vector construction, and protein prokaryotic function expression. Background technique [0002] When it comes to tuberculosis, everyone's eyes turn pale. That's why, let's first look at a set of data. Judging from the situation announced by the World Health Organization, tuberculosis has become one of the most serious diseases threatening human health. my country is one of the 22 countries with a high burden of tuberculosis in the world, and about 550 million people in my country are infected with tuberculosis. It can be seen that it is particularly important to explore the infection mechanism of tuberculosis. Mycobacterium tuberculosis is the caus...

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Application Information

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IPC IPC(8): C07K14/35C12N15/70C12N15/10C12N1/21C12R1/19
CPCC07K14/35C12N15/70
Inventor 袁建波黎兵向加林胡姗姗黄健陈泽慧闵迅
Owner AFFILIATED HOSPITAL OF ZUNYI MEDICAL COLLEGE
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