RECOMBINANT PLASMID CONTAINING 3'-TERMINUS CDNA FRAGMENT OF INFLUENZA-A VIRUS NP-GENE CONSTRUCTION AND EXPRESSION IN ESCHERICHIA-COLI-CELLS

A recombinant plasmid pN62 determining the synthesis of a hybrid protein consisting of a full-size beta-galactosidase and C-terminus fragment of influenza A virus nucleoprotein was constructed. The complete identity oi pN62 insert with the 3'-terminus cDNA fragment of influenza A virus NP gene and conservation of beta-galactosidase reading frame was confirmed by Maxam-Gilbert sequencing of pN62. An expression of pN62 plasmid in E. coli JM103 in the presence of IPTG resulted in accumulation of fused protein as poorly soluble inclusion bodies in the bacterial cells. Analysis of E. coli JM103/pN62 bacteria lysates by 7 % PAAGE revealed that molecular weight of hybrid polypeptide was 18 kDa heavier than normal beta-galactosidase and corresponded to the previously deduced weight of 135 kDa.