Diblock fusion protein with adhesion-antifreeze dual functions, and synthesis method and application thereof

Abstract

The invention relates to a diblock fusion protein with adhesion-antifreeze dual functions, and a synthesis method and application thereof. By taking genetic engineering as a technological means, a gene expression vector simultaneously containing antifreeze protein genes and adhesion protein genes is prepared and converted into a host cell, through an inducer, the host cell overexpresses the protein, and thus the fusion protein is obtained by separation and purification. The amino acid sequence of the fusion protein is SEQ ID NO.3, and the nucleotide sequence of the fusion protein is SEQ ID NO.4. The mussel adhesion protein and the antifreeze protein are fused on the gene level, and thus the diblock fusion protein with the adhesion-antifreeze dual functions is constructed; and the fusion protein is expressed through fermentation of escherichia coli, supernatant protein is purified through an affinity chromatogram, an inclusion body is purified through acetic acid, a high-purity separation and purification method is constructed, the fusion protein is synthesized through a microbial one-step method, the process is mild and efficient, the cost is low, the equipment requirement degree is low, and expanded production at the later period is easy.

Description

technical field

[0001] The invention belongs to the field of biotechnology, and specifically relates to a diblock fusion protein with dual functions of adhesion and antifreeze, its synthesis method and application; including the gene design of the diblock fusion protein with dual functions of adhesion and antifreeze, Construction of recombinant expression vector containing fusion protein gene fragment, fermentation expression in Escherichia coli, separation and purification method of fusion protein, and application on antifreeze surface. Background technique

[0002] Antifreeze protein (AFP) is a general term for a class of protein compounds that can improve biological antifreeze ability. It can bind to small ice crystals and prevent ice crystallization and crystal growth. Generally, the larger the thermal hysteresis value of the antifreeze protein, the better the adaptability of the organism to low temperature. (The thermal hysteresis effect is to reduce the freezing point...

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