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

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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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: Disadvantages: slow growth, high nutritional requirements, it takes more than 10 hours to cultivate one generation; the cell wall is thick and rich in lipids, which hinders the molecular transportation including exogenous DNA; the transformation efficiency is low; the expression of exogenous genes requires special vectors, etc.

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