Methods and compositions for secretion of heterologous polypeptides
First Claim
1. A method of secreting a heterologous polypeptide of interest in a cell comprising using a translational initiation region variant operably linked to nucleic acid encoding said heterologous polypeptide to express said heterologous polypeptide, wherein the translational strength of said variant translational initiation region is less than the translational strength of the wild-type translational initiation region and wherein the amino acid sequence of said translational initiation region variant is not altered.
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Abstract
The instant invention discloses the unexpected result that mutant signal sequences with reduced translational strength provided essentially complete processing and high levels of expression of a polypeptide of interest as compared to wild type signal sequences, and that many mammalian polypeptides require a narrow range of translation levels to achieve maximum secretion. A set of signal sequence vectors provides a range of translational strengths for optimizing expression of a polypeptide of interest.
279 Citations
21 Claims
- 1. A method of secreting a heterologous polypeptide of interest in a cell comprising using a translational initiation region variant operably linked to nucleic acid encoding said heterologous polypeptide to express said heterologous polypeptide, wherein the translational strength of said variant translational initiation region is less than the translational strength of the wild-type translational initiation region and wherein the amino acid sequence of said translational initiation region variant is not altered.
- 10. A nucleic acid encoding a translational initiation region variant, wherein the translational strength of said variant translational initiation region is less than the translational strength of the wild-type translational initiation region, wherein said translation initiation region includes a prokaryotic secretion signal sequence selected from the group consisting of OmpA, PhoE, MBP and PhoA.
- 11. A nucleic acid encoding a polypeptide operably linked to a translational initiation region variant, wherein the translational strength of said variant translational initiation region is less than the translational strength of the wild-type translational initiation region, wherein the amino acid sequence of said translational initiation region variant is not altered.
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21. A method of secreting a heterologous polypeptide of interest in a cell comprising using a translational initiation region variant operably linked to nucleic acid encoding said heterologous polypeptide to express said heterologous polypeptide, wherein the translational strength of said variant translational initiation region is less than the translational strength of the wild-type translational initiation region wherein said translation initiation region includes a prokaryotic secretion signal sequence selected from the group consisting of OmpA, PhoE, MBP and PhoA.
Specification