Procaryotic leader sequence in recombinant baculovirus expression system
AssigneeBurroughs Wellcome Co.
InventorsTimothy C. Peakman, Martin J. Page, Ian G. Charles
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Abstract
A process for the preparation of a protein, which process comprises culturing under such conditions that the protein is obtained cells infected with a recombinant baculovirus having a promoter, capable of directing the expression of a gene in the cells, operably linked to a heterologous gene which encodes the protein and which is preceded by a leader sequence comprising: T A A T C A T C C A C A G G A G A C T T T C T G [SEQ. ID NO: 1].
Claims20
A process for preparing a protein, which process comprises culturing, under such conditions that said protein is obtained, insect cells infected with a recombinant baculovirus having a promoter capable of directing expression of a heterologous gene having a coding sequence encoding said protein, said promoter being operably linked to said heterologous gene and said coding sequence being preceded by an untranslated leader of sequence
- T A A T C A T C C A C A G G A G A C T T T C T G (SEQ ID No:
- 1).
A recombinant baculovirus having a promoter capable of directing expression of a heterologous gene having a coding sequence in insect cells infected with said recombinant baculovirus, said promoter being operably linked to said heterologous gene, and said coding sequence being preceded by an untranslated leader of sequence
- T A A T C A T C C A C A G G A G A C T T T C T G (SEQ ID No:
- 1).
A process for preparing a recombinant baculovirus, which process comprises:
- (a) providing a recombinant baculovirus transfer vector having a promoter capable of directing expression of a heterologous gene in insect cells infected with a recombinant baculovirus, said promoter being operably linked to said heterologous gene, and said coding sequence being preceded by an untranslated leader of sequence
- T A A T C A T C C A C A G G A G A C T T T C T G (SEQ ID No:
- 1);
- and
- (b) co-transfecting said cells with said vector and intact wild type baculovirus DNA.
A recombinant baculovirus transfer vector for use in the preparation of a recombinant baculovirus, which vector has a promoter capable of directing expression of a heterologous gene having a coding sequence in insect cells infected with said recombinant baculovirus, said promoter being operably linked to said heterologous gene, and said coding sequence being preceded by an untranslated leader of sequence
- T A A T C A T C C A C A G G A G A C T T T C T G (SEQ ID No:
- 1).
A baculovirus transfer vector comprising
- (i) a restriction site into which a heterologous gene may be cloned downstream of the sequence encoding the N-terminus of polyhedrin;
- (ii) another triplet of nucleotides in place of the natural ATG translation start condon of the polyhedrin coding sequence;
- (iii) from up to the first 24 to up to the first 50 bases of the polyhedrin coding sequence;
- and
- (iv) downstream of (iii) but upstream of (i), an untranslated leader of sequence
- T A A T C A T C C A C A G G A G A C T T T C T G (SEQ ID NO:
- 1).
Dependencies are inferred from the claim text.