Recombinant expression vector of Hsp65-hIL-2 fusion expression and recombinant strain

A technology that combines expression and expression vectors, applied in the biological field, can solve problems such as high nutritional requirements, slow growth, and cell wall thickness, and achieve the effect of improving killing activity

Inactive Publication Date: 2011-09-07
FOURTH MILITARY MEDICAL UNIVERSITY
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Problems solved by technology

[0006] Introducing exogenous genes into mycobacteria to construct recombinant mycobacterial live vaccines is one of the important directions of new TB vaccine research. Most domestic and foreign studies use BCG as the carrier of mycobacterial live vaccines, but recombinant BCG vaccines still exist as follows: Disadvan

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  • Recombinant expression vector of Hsp65-hIL-2 fusion expression and recombinant strain
  • Recombinant expression vector of Hsp65-hIL-2 fusion expression and recombinant strain
  • Recombinant expression vector of Hsp65-hIL-2 fusion expression and recombinant strain

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Embodiment Construction

[0036] The present invention provides an expression vector pDE-hsp65-hIL-2 expressing the fusion expression of Hsp65-hIL-2, and transfecting it into Mycobacterium smegmatis to prepare recombinant MS, and obtaining Mycobacterium smegmatis Hsp65 through hygromycin resistance screening -hIL-2-MS; and verify the expression of the target gene and the immune effect of Hsp65-hIL-2-MS, the results show that the recombinant Hsp65-hIL-2-MS is effective in preparations or medicines for the prevention and treatment of tuberculosis The application has a good application prospect. The present invention will be further described in detail below in conjunction with the accompanying drawings, which are explanations rather than limitations of the present invention.

[0037] 1. Construction of pDE-hsp65-hIL-2 Escherichia coli-mycobacterium shuttle secretion expression vector expressed by Hsp65-hIL-2 fusion

[0038] 1.1 Cloning of HSP65 protein gene

[0039] Primers were designed according to t...

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Abstract

The invention discloses a recombinant expression vector of Hsp65-hIL-2 fusion expression. An Hsp65-hIL-2 fusion gene is inserted into an escherichia coli-mycobacterium shuttle secreting expression vector, and the Hsp65-hIL-2 fusion gene is formed by connecting an Hsp65 gene and hIL-2 through an enzyme cutting site and a linker. According to a recombinant strain of Hsp65-hIL-2 fusion expression, mycobacterium smegmatis is used as a host cell, and an exogenous expression vector for transforming the host cell is the escherichia coli-mycobacterium shuttle secreting expression vector containing the Hsp65-hIL-2 fusion gene. The mycobacterium smegmatis Hsp65-hIL-2-MS recombinant strain centralizes the advantages of a target antigen, a cytokine adjuvant and a live vector; and the immune inoculated recombinant strain can stimulate the body to produce immune response comprising body fluid immunity level and cellular immunity level and improve the immune protection of the body.

Description

technical field [0001] The invention belongs to the field of biotechnology and relates to the development of a new TB vaccine, in particular to a recombinant expression vector and a recombinant bacterial strain for fusion expression of Hsp65-hIL-2. Background technique [0002] According to the WHO report, about 2 billion people in the world have been infected with Mycobacterium tuberculosis (MTB), of which 50 million may be infected by drug-resistant strains, about 10 million new patients are diagnosed each year, and the annual death toll is as high as 3 million. Among all infectious diseases, the death rate of tuberculosis (TB) is second only to AIDS. WHO lists TB, together with AIDS and malaria, as the most important infectious killer of human beings. my country is one of the 22 countries with a high burden of TB in the world. The number of deaths due to TB is about 250,000 each year. It ranks first among the 27 legally notifiable infectious diseases in my country and is...

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Application Information

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IPC IPC(8): C12N15/74C12N1/21A61P31/06C12R1/34
Inventor 徐志凯王丽梅柏银兰师长宏张海康健王平张薇
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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