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A kind of recombinant Periplaneta americana thymosin protein pathy1 and its expression method

A technology of American cockroach and thymosin, applied in the field of genetic engineering, can solve the problems of difficult purification and low thymosin content

Active Publication Date: 2021-04-20
SICHUAN GOODDOCTOR PANXI PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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

Method used

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  • A kind of recombinant Periplaneta americana thymosin protein pathy1 and its expression method
  • A kind of recombinant Periplaneta americana thymosin protein pathy1 and its expression method
  • A kind of recombinant Periplaneta americana thymosin protein pathy1 and its expression method

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Embodiment 1

[0029]Test material

[0030]1.1 America big sample

[0031]The American Damu sample is provided by the Sichuan Nice Doctor Panxi Pharmaceutical Co., Ltd. Xichang Farm Base.

[0032]1.2 plasmid vector and receptor strain

[0033]The gene prokaryotic expression vector PET28A is preserved for this laboratory. Escherichia coli DH5A, raw nuclear expression cells BL21 (DE3) were purchased from Bourway Co., Ltd.

[0034]1.3 restriction endonuclease and other enzymes

[0035]Restriction endonuclease (XHOI, ECOR I), T4DNA Ligase (DNA ligase), TAQ enzymes are available in Chengdu Tiantai (Takara) Bio Co., Ltd.

[0036]1.4 instrument equipment and consumables

[0037]UV spectrophotometer, PCR amplifier (Biometra, Germany), vertical pressure steam sterilization pot (Shanghai Boxing Industrial Co., Ltd.), protein vertical electric soot tank, transfer electrophoresis tank, shaker, oscillator, ultrasonic crusher, protein Purification instrument.

[0038]1.5 medium and solution

[0039]1.5.1 Liquid LB medium

[0040]100ml formula:...

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Abstract

The invention relates to the field of biotechnology, and discloses a recombinant Periplaneta americana thymosin protein PaTHY1 and an expression method thereof. The method comprises the following steps: (1) extracting RNA from Periplaneta americana tissue, and reverse transcribing it into cDNA; (2) Synthetic primers: (3) using the total cDNA as a template to obtain a thymosin protein coding gene fragment of Periplaneta americana by PCR; (4) cloning the coding gene into a prokaryotic expression vector, transforming the host bacteria, culturing the obtained recombinant engineering bacteria, and then inducing The target protein is expressed; (5) the target protein is purified by affinity chromatography, thereby obtaining the recombinant Periplaneta americana thymosin protein PaTHY1. The recombinant protein was obtained through genetic engineering for the first time, solved the problem of extremely low content of thymosin in the complex chemical components of American cockroach and difficult purification, and laid a foundation for the research on the pharmacodynamics of thymosin in American cockroach.

Description

Technical field[0001]The present invention relates to the field of genetic engineering, and in particular, there is a recombinant American thymine protein pATHY1 and its expression methods.Background technique[0002]Thymosin (Thy) is mainly a lymphocyte growth factor produced by thymus, which is generally present in various tissue cells. First, Goldstein and White were first found in 1966 from the fetal boar thymom protein extract, a group of small molecular polypeptides, containing more than 40 components, thus being named thymoxin. The thymoxin can be divided into α, β, γ3 family according to the equidum (PI), of which Pi is called β-Thymosin, Tβ), which is highly conserved by 40- The 44 amino acids composition, the molecular mass is about 5.0ku, and is widely present in vertebrate and invertebrates. The β-stranded thymocarin is a polar protein molecular family, which was initially considered to be a thymine hormone, which had different degrees of characteristics that affect blood ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/575C12N15/70C12R1/19
CPCC07K14/57581C12N15/70C12N2800/101
Inventor 耿福能岳碧松沈咏梅
Owner SICHUAN GOODDOCTOR PANXI PHARMA
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