RECOMBINANT PLASMID DNA PET32M/MTRX-RHIL-11, ENCODING INTERLEUKIN-11 OF MAN, METHOD FOR ITS OBTAINMENT AND STRAIN ESCHERICHIA COLI BL21 (DE3) /PET32M/MTRX-RHIL-11 - PRODUCER OF RECOMBINANT INTERLEUKIN -11

摘要

1. Recombinant DNA plasmid pET32M / mTrx-rhIL-11 coding for human IL-11 as a part of fused protein with thioredoxin I E.coli, consisting of rET32M plasmid and nucleotide sequence of human IL-11 synthetic gene comprising a promoter T7lac, altered gene Asn84 / Gln thioredoxin I E.coli, hexa-histidine cluster site Asn-Gly, synthetic gene of human IL-11, T7 terminator, the ampicillin resistance gene bla gene and lactose operon repressor lacI, designed for expression in E.coli cells. ! 2. A method for producing a recombinant plasmid DNA pET32M / mTrx-rhIL-11, comprising the synthesis of four gene fragments of recombinant IL-11 of oligonucleotide primers in the cloning of each fragment digested with endonuclease EcoRV plasmid vector pGEM5z, transforming each of the four plasmids E.coli strain DH5α cells , selection of the appropriate gene fragments encoding IL-11 clones selected plasmids with the fragments of IL-11 gene without mutations, the union of all the fragments of the gene recombinant human IL-11 into the plasmid vectors D pUC19, construction rET32M vector based on plasmid pET32b, pereklonirovanie gene recombinant IL-11 in rET32M expression vector carrying a gene mutant thioredoxin (mTrx). ! 3. The strain of E.coli BL21 (DE3) / FP, transformed with the recombinant plasmid DNA pET32M / mTrx-rhIL-11 - Producing recombinant human interleukin-11 consisting of a soluble protein fused mTrx-rhIL-11.