Baculovirus expression system capable of producing foreign gene proteins at high levels

US5194376A · kind A · utility

Assignee

• THE UNIVERSITY OF OTTAWA · CA

Inventor

• C. Yong Kang · Ottawa, CA

Key dates

• Filing date: Feb 28, 1989
• Grant date: Mar 16, 1993
• Expiry date: Feb 28, 2009

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12N2740/16322
• WIPO field: Pharmaceuticals
• WIPO sector: Chemistry

Abstract

A baculovirus expression system capable of producing foreign gene proteins at high levels. The system involves the production of a recombinant baculovirus containing a modified foreign gene between the polyhedrin gene promoter region and the transcription termination signal of the polyhedrin structural gene. The modified foreign gene comprises a putative ribosome binding site immediately upstream of the foreign gene coding sequence, i.e. without any intervening non-coding sequences. The putative ribosome binding site is preferably properly positioned without the intervening sequences by a crossover linker mutagenesis procedure before the modified foreign gene is introduced into the virus. The putative ribosome binding site preferably comprises at least the final four nucleotides of the sequences ACCTATAAAT immediately upstream of the translation inibiation codon (ATG) of the foreign gene. The system is capable of producing foreign gene proteins (when insect cells are infected with the recombinant virus) at high levels, even in the case of those genes which expressed only at low or intermediate levels in prior recombinant baculovirus systems.