Baculovirus expression system capable of producing foreign gene

Details Baculovirus expression system capable of producing foreign gene Baculovirus expression system capable of producing foreign gene

1989-02-28

1993-03-16

Schwartz, Richard A.

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Recombinant dna technique included in method of making a...

4351723, 4353201, C12N 1500, C12N 1586, C12P 2102

Patent

active

051943760

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.

REFERENCES:
patent: 4745051 (1988-05-01), Smith et al.
C. Yong Kang, "Baculovirus Vectors for Expression of Foreign Genes"--Advances in Virus Research, vol. 35 pp. 177-192.
Matsuura, Y. et al.--"Baculovirus Expression . . . "--J. Gen. Virol. (1987), 68, 1233-1250.
Horiuchi, T. et al.--"High-Level Expression of the Human-.alpha.-Interferon Gene . . . "--Ag. Biol. Chem. 51(6), 1573-1580, 1987.
Kozak, M.--"Point Mutations Define A Sequence Flanking The AUG Initiator Codon . . . "--Cell, vol. 44, 283-292, Jan. 31, 1986.
Sung, W. L.; et al.--"Synthesis of Mutant Parathyroid Hormone Genes via Site-Specific . . . "--Gene, 47 (1986) 261-267.
Maeda, S. et al.--"Production of Human .alpha.-Interferon in Silkworm Using a Baculovirus Vector"--Nature vol. 315, Jun. 13, 1985.

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