Of recombinant human thymosin β 4 gene cloning expression purification identification and activity detection.docVIP

 PAGE \* MERGEFORMAT 10 Of recombinant human thymosin β 4 gene cloning expression purification identification and activity detection [Abstract] Objective: Using genetic engineering of prokaryotic expression of recombinant human thymosin β 4 (Tβ 4), and purification and identification of them. Methods: The use of synthetic human codon preference of E. coli Tβ 4 full-length gene was constructed Tβ 4 body of the second series gene (Tβ 4 ② ), and cloned into the expression vector pET  22b (), the recombinant plasmid was transformed into E. coli BL21 (DE3) Competent, IPTG induced expression, via salt precipitation, hydrophobic chromatography and ion exchange chromatography purification of recombinant protein , using Western blot and matrix-assisted laser desorption ionization time of flight mass spectrometry of the recombinant proteins were identified. Results: Tβ 4 ② of the purified recombinant protein, and has biological activity. Conclusion: The successful construction, expression and purification of the Tβ 4 ② , its Function lay the foundation. [Keywords:] Thymosin β 4 tandem repeat sequence of molecular cloning of gene expression 0 Introduction Thymosin β 4 (thymosin beta 4, Tβ 4) widely distributed in various tissues, organs and cells, and immune function, neurological development and function of actin in close, but also anti-inflammatory, to promote angiogenesis [1], wound healing [2 ], hair follicle development [3], repair damaged heart muscle [4]. Although Tβ 4 have broad application prospects, but because of its small molecule, difficult to express directly in E. coli. currently used in laboratory and clinical trials Tβ 4 mainly synthetic materials, is expensive, its use is restricted. We use gene synthesis and PCR technology, the two series was constructed Tβ 4 gene (Tβ 4 ② ) expression vector and successfully expressed in E. coli and purification of the body series , in order to lay a foundation for further research. 1 Materials and methods 1.