Recombinant thymosin protein PaTHY1 of Periplaneta americana and expression method thereof

Abstract

The invention discloses a recombinant thymosin protein PaTHY1 of Periplaneta americana and an expression method thereof, belonging to the field of biotechnology. The expression method comprises the following steps: (1) extracting RNAs from the tissue of Periplaneta americana and carrying out inverse transcription on the RNAs to obtain cDNAs; (2) synthesizing primers; (3) with the cDNAs as a template, carrying out PCR to obtain a coding gene fragment of the thymosin protein of Periplaneta americana; (4) cloning the coding gene fragment into a prokaryotic expression vector for transformation ofhost bacterium, carrying out culturing on an obtained recombinant engineering bacterium and inducing the recombinant engineering bacterium to express the target protein; and (5) purifying the target protein by using affinity chromatography so as to obtain the recombinant thymosin protein PaTHY1 of Periplaneta americana. The recombinant protein is obtained through genetic engineering for the firsttime, overcomes the problems of low content and difficult purification of thymosin in complex chemical components of Periplaneta americana, and lays a foundation for research on the pharmacodynamic functions of thymosin from Periplaneta americana.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to a recombinant Periplaneta americana thymosin protein PaTHY1 and an expression method thereof. Background technique [0002] Thymosin (THY) is mainly a lymphocyte growth factor produced by the thymus, which is ubiquitous in various tissue cells. It was first discovered by Goldstein and White in 1966 from the extract of fetal bovine thymus protein. It is a group of small molecular polypeptides containing more than 40 components, so it is named thymosin. According to the isoelectric point (PI), thymosin can be divided into three families: α, β, and γ. Among them, the PI of 5.0-7.0 is called β-thymosin (β-thymosin, Tβ), and its structure is highly conserved. It consists of 40- Composed of 44 amino acids, the molecular mass is about 5.0ku, and widely exists in vertebrates and invertebrates. β-thymosin belongs to the family of polar protein molecules, and was originally considered ...

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Problems solved by technology

[0005] The object of the present invention is to provide a kind of Periplaneta americana thymosin protein PaTHY1 through genetic engineering, and provide a kind of expression method of American cockroach thymosin protein PaTHY1, solve the extremely low content of thymosin in the American cockroach extract, purify difficulty problems, and lay the foundation for the study of thymosin function in Periplaneta americana

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