Recombinant plasmid DNA Per-TB4GyrA-AcSer, encoding the serine acetyltransferase, the ability of IN VIVO acetylated N-terminal serine Desacetylthymosin BETA 4 and the hybrid protein capable of autocatalytic cleavage with formation Thymosin Beta 4 HUMAN STRAIN Eschrichia coli C3030 / pER-TB4GyRA-AcSer PRODUCER These proteins and methods for producing genetically engineered thymosin Beta 4 HUMAN

摘要

1. A recombinant plasmid DNA pER - TB4GyrA - AcSer, containing polycistronic structure for simultaneous biosynthesis of two proteins - a fusion polypeptide containing thymosin β4 man and intein, and the enzyme of E.coli - serine acetyltransferase in cells of Escherichia col; NdeI / SapI - a fragment of the plasmid DNA pTWIN-1; NdeI / SapI - DNA fragment containing adapted to these sites gene sequence of recombinant human thymosin β4 and comprising, as a genetic marker β-lactamase gene that determines the stability of the transformed plasmid pER - TB4GyrA - AcSer E.coli cells to penicillin antibiotics; unique restriction endonuclease recognition sites located at a distance next to the left from the site BamHI - Pstl - 575 bp, NdeI -: 1493 bp, XbaI - 1532 bp, EcoRV - 3565 bp, HpaI - 3621 p.o.2. Escherichia coli Strain S3030 / pER - TB4GyrA - AcSer, producing a fusion polypeptide comprising a human β4 Desacetylthymosin and intein and serine acetyltransferase obtained by transforming Escherichia coli cells of the strain S3030 recombinant DNA plasmid pER - TB4GyrA - AcSer of claim 1.3.. A method for producing recombinant thymosin β4 human comprising transformation of strain Escherichia coli S3030 plasmid DNA pER - TB4GyrA -. AcSer, according to claim 1, culturing the producing strain Escherichia coli S3030 / pER - TB4GyrA -. AcSer according to claim 2, separation TB4GyrA fusion protein in the soluble form following cell disruption using ultrasonic disintegrator in a buffer solution (50 mM Tris / HCl, 10 mM EDTA, 1 mM PMSF), adsorbing the fusion protein to kolonkes chitinous sorbent (NEB, England), inducing its autocatalytic cleavage washing chitin sorbent buffer solution containing balanced in a buffer containing