Recombined extrasin alpha 1 two-strand body protein and preparation method thereof

A technology of thymosin and protein, which is applied in the field of thymosin α1 dimer protein and its preparation, can solve the problems affecting the application of recombinant protein, the purification process is complicated, and it is not suitable for industrial production, so as to achieve easy scale-up production and simple separation and purification process Effect

Inactive Publication Date: 2009-09-30
FOURTH MILITARY MEDICAL UNIVERSITY
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Problems solved by technology

[0031] Although the above methods can obtain active recombinant proteins, there are still many shortcomings. The process of obtaining thymosin α1 monomer by chemical cleavage is often very complicated in the later purification process; the recombinant prot

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  • Recombined extrasin alpha 1 two-strand body protein and preparation method thereof
  • Recombined extrasin alpha 1 two-strand body protein and preparation method thereof
  • Recombined extrasin alpha 1 two-strand body protein and preparation method thereof

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Embodiment Construction

[0068] The technical scheme adopted in the present invention is: to construct pET-22b(+)-Tα1② prokaryotic expression vector, to efficiently express thymosin α1 in E. The activity shows that the thymosin α1 dimer protein with biological activity is isolated, and the separation and purification process is simple, cheap and easy to expand production. In addition, in vitro experiments have proved that the purified thymosin α1 dyad can inhibit the proliferation of HL-60, B16, S180, HepG-2, M1 and other tumor cells; the purified thymosin α1 dyad intervened in B16 tumor-bearing mice, Can significantly inhibit tumor growth, and its effect is better than chemical synthesis.

[0069] The above scheme is specifically carried out according to the following steps:

[0070] 1) Construct thymosin α1 dyad gene expression vector or recombinant plasmid pET-22b(+)-Tα1②

[0071] a) Design and synthesis of thymosin distring gene (Tα1②)

[0072] According to the codon preferred by Escherichia co...

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Abstract

The invention relates to a recombined extrasin alpha 1 two-strand body protein and a preparation method thereof. Glycin and serine are connected in series with bimolecular extrasin alpha 1 (T alpha 1), a connection mode is as follows: T alpha 1-Gly-Ser=T alpha 1, and an extrasin alpha 1 two-strand body gene expression vector is constructed by synthesized extrasin alpha 1 two-strand body genes so that extrasin alpha 1 which can not be directly expressed in colon bacillus can be efficiently expressed in the colon bacillus in a two-strand body way. The purified extrasin alpha 1 two-strand body protein has biological activity and can promote lymphocytes of small mice to proliferate, restrain tumor cell strains from proliferating and obviously restrain the tumor growth of tumor-bearing mice, thereby establishing a base for the further research and the extensive application of the extrasin alpha 1.

Description

technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to the construction, prokaryotic expression, purification and identification of recombinant plasmids of target proteins, in particular to thymosin alpha 1 (T alpha 1) dimer protein and its preparation method. Background technique [0002] 1. Biochemical properties of thymosin α1 [0003] 1.1 Thymosin family [0004] Thymosin is mainly produced by the thymus. It was originally prepared by Goldstein from the fetal bovine thymus protein extract. It contains more than 40 polypeptide components. According to the position of these polypeptides on the isoelectric focusing electrophoresis analysis map, they are divided into α, β, and γ. Three regions: α region pI<5, β region pI between 5 and 7, and γ region pI>7. The α-thymosin group includes thymosin α, prothymosin α and parathymosin α. Thymosin α1 (Tα1) is an N-terminal 28 amino acid fragment of prethymosin α that is produ...

Claims

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Application Information

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IPC IPC(8): C07K14/66C12N15/16C12N15/70C07K1/20C07K1/18A61K38/32A61P35/00A61P37/04A61P31/12A61P7/06
Inventor 张英起韩苇李维娜
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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