RECOMBINANT PLASMID DNA PHBMD1 CONTAINING THE MODIFIED GENE OF MEDIAN SURFACE PROTEIN OF HUMAN HEPATITIS VIRUS B, - INTERMEDIATE PLASMID FOR RECOMBINANT PLASMID DNA YEP 63/AB CONSTRUCTING, RECOMBINANT PLASMID DNA YEP 63/AB ENCODING SYNTHESIS OF MEDIAN SURFACE DERIVATIVE PROTEIN OF HEPATITIS B VIRUS AND STRAIN OF YEAST SACCHAROMYCES CEREVISIAE - A PRODUCER OF MEDIAN SURFACE DERIVATIVE PROTEIN OF HEPATITIS B VIRUS

摘要

FIELD: molecular biology, genetic engineering, medicine. SUBSTANCE: recombinant plasmid pHBmd1 that is inherited in E. coli cells and carrying the modified gene of median surface protein of human hepatitis B virus (M-protein HBV) and containing deletion in specific site of region preS2 that determines sensitivity of M-protein HBV to trypsin-like proteases is obtained on the basis of the cloned gene encoding M-protein HBV. On the basis of plasmid pHBmd1 recombinant plasmid YEp63/AB is constructed. This plasmid provides synthesis of M-protein HBV derivative in the transformed cells of yeast S. cerevisiae, strain 20B12. The presence of intact promoter of yeast gene encoding glyceraldehyde-3-phosphate dehydrogenase in plasmid YEp63/AB located before initiating codon AUG ensures to get high expression level without induction. Synthesized protein assembles particle normally as the natural 22 nm particle HBsAg. Synthesis level of M-protein derivative is 0.5-1% of total protein level in cell. New strain-producer can be used for industrial production of M-protein HBV derivative and used for diagnostic and immunoprophylaxis aims, for development of antihepatitis vaccines of new generation showing the enhanced protective properties. EFFECT: increased proteolytic stability of M-protein HBV derivative, new strain indicated above.