RECOMBINANT PLASMID DNA PESG ENCODING FUSED PROTEIN SYNTHESIS THAT SHOWS ANTIGENIC PROPERTY OF HUMAN T-CELLULAR LEUKOSIS I-TYPE VIRUS SURFACE GLYCOPROTEID ENVELOPE (HTLV-I), METHOD OF ITS CONSTRUCTION AND STRAIN OF BACTERIUM ESCHERICHIA COLI HB 101/PESG - PRODUCER OF FUSED PROTEIN

摘要

FIELD: genetic engineering, biotechnology. SUBSTANCE: recombinant plasmid DNA PESG (size is 5.6 thousand nucleotide pairs) consists of SalGI-BamHI-fragment (426 nucleotide pairs) of region sequence env HTLV-I and SalGI-fragment of plasmid pUR291 (size is 5.2 thousand nucleotide pairs) with beta-galactosidase gene, region ori and genetic marker as bla-gene (resistance to ampicillin) and has the unique restriction sites to BamHI, PstI, HindIII, StuI, ClaI, Tth111-II. Fragment (size is 426 nucleotide pairs) is excised from intermediate plasmid pBE1 using restriction enzyme SalGI and this fragment is ligated with SalGI-fragment of plasmid vector DNA pUR291 linearized by hydrolysis. After incubation competence cells of E. coli (treated with CaCl₂) were transformed with obtained preparation followed by selection of transformants according their capability to grow on ampicillin-containing medium. Proposed invention ensures to obtain 400-500 mg fused protein from 1 l cell suspension of E. coli that contains specific polypeptide sequence encoded by region env of genome HTLV-I. EFFECT: construction of recombinant plasmid DNA, high level of fused protein production, construction of the strain-producer indicated above.