Construction of lactobacillus acidophilus S-layer protein surface display system

Abstract

The invention provides construction of a lactobacillus acidophilus S-layer protein surface display system, which belongs to the field of biotechnology. The construction is characterized in that gene acquisition comprises (1) PCR amplification, (2) the connection of a SlpA gene and a pMD18-TVector system and (3) the identification of cloning plasmid pMD18T-S, and the construction comprises (1) theacquisition of a target gene and a double-labeling expression vector, (2) the connection and transformation of the double-labeling expression vector and SlpA, (3) the identification of a surface display vector system and (4) the detection of the anchored expression situation of S-layer protein on recombinant surface. The construction has the advantages that: 1, the constructed vector can be used to be transformed into S-layer protein gene defect lactobacillus to study the formation mechanism of lactobacillus S-layer protein; and 2, by utilizing a DNA recombination technique, protective antigengenes of pathogens are fused with lactobacillus S-layer protein on the surface display vector.

Description

technical field [0001] The invention belongs to the field of biotechnology. Background technique [0002] 1 Microbial cell surface display technology [0003] Microbial cell-surface display (MCSD) refers to the use of techniques such as molecular biology and immunology to anchor and display exogenous protective peptide molecules and protein molecules on the surface of microbial cells. The former is fused with a certain surface protein of the cell, and the former is positioned on the cell surface by means of the surface recognition and surface positioning functions of the bacterial protein. The former is called the passenger protein, and the latter is called the carrier protein. The expressed gene product does not need to be extracted. , purification, renaturation and other operations, you can directly use the whole cell to carry out the follow-up work of expressing the protein [1] . [0004] 2 Research Status of Surface Display System [0005] 2.1 Phage surface display s...

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Problems solved by technology

The slpC gene was isolated from the slpB gene. Sequence analysis showed that the slpC gene has a complete promoter and terminator, and has the ability to encode SlpC, and the slpC gene can be detected from the bacterial genome under aerobic and anaerobic conditions. It exists, but there is no expression of related proteins. It is speculated that the product of the slpB gene may limit the expression of the slpC gene

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