Method for preparing recombinant human keratinocyte growth factor-2 by secretion expression

Abstract

The invention provides a method for preparing recombinant human keratinocyte growth factor-2 by secretion expression. The method provides an expression vector of the recombinant human keratinocyte growth factor-2, which contains an secretion signal peptide coding sequence and can secrete KGF-2 protein in periplasm for expression; and when a host cell containing the recombinant plasmid is subjected to repeated freeze thawing at a low temperature, the recombined protein can be released in a buffer solution under the condition of not damaging inner membranes of bacilli and the activity of the protein.

Description

A preparation method for secreting and expressing recombinant human keratinocyte growth factor-2 technical field The invention relates to a preparation method for optimally secreting and expressing recombinant human keratinocyte growth factor (rhKGF-2), as well as an expression vector and a host cell used in the method. Background technique Keratinocyte growth factor (keratinocyte growth factor, KGF) was isolated and purified from the culture supernatant of human fetal lung fibroblasts by Rubin et al. in 1989, and was found to have the effect of promoting mouse keratinocyte mitosis, so it was named Keratinocyte growth factor. In 1996, Yamasaki et al. amplified a new growth factor from mouse embryos, another new cytokine closely related to KGF, called KGF-2, also known as fibroblast growth factor-10 (FGF -10). Afterwards, Emoto et al. cloned the cDNA of human KGF-2 from human lungs. The gene is located at 5p12-5p13 of human chromosome 5. Human FGF-10 consists of 624 nucle...

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Problems solved by technology

At present, the production process of the rhKGF-2 E. coli expression system in the world, whether the expression product exists in the form of soluble protein or not, is expressed in the cytoplasm, and the protein must be broken by ultrasonic waves to release the protein, which not only affects the protein itself Has a destructive effect, but also increases the difficulty and cost of purification

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