DNA FRAGMENT ENCODING PROTEIN GELONIN AND METHOD OF ITS PRODUCING, VECTOR PROVIDING PROTEIN GELONIN EXPRESSION, RECOMBINANT STRAIN ESCHERICHIA COLI - PRODUCER OF PROTEIN GELONIN

摘要

FIELD: biotechnology, molecular biology, genetic engineering. SUBSTANCE: synthetic gene encoding protein gelonin is obtained. This nucleotide sequence SEQ 1D N1 is determined and given in the invention description. Method of producing involves the stage of selection of DNA fragment sequence, the stage of synthesis that includes the separation of DNA fragment for two overlapping sets of single-stranded fragments of oligonucleyides. Chemically synthesized oligonucleotides are taken among the group consisting of Btgel 1, Btgel 2 and gel 1-gel 39 followed by their purification and phosphorylation at 5'-end. Annealing of oligonucleotide strands in combination with DNA duplex formation results to the formation of the synthetic gene. Also, recombinant strain E. coli transformed by the expression vector that provides the expression of protein gelonin is obtained. Vector has DNA fragment showing the sequence SEQ ID NI ligated in Eco RI/Hind III-fragment split by vector 223-3. This recombinant technology ensures to obtain the rich and reproducing source of gelonin that is an immunotoxic agent used for treatment of tumors. EFFECT: improved method of producing, valuable antitumor properties of the end product. 5 cl, 5 dwg, 9 ex