Modified MSP-1 nucleic acid sequences and methods for increasing MRNA levels and protein expression in cell systems
First Claim
Patent Images
1. A modified nucleic acid sequence comprising SEQ ID NO 1.
6 Assignments
0 Petitions
Accused Products
Abstract
The invention provides modified recombinant nucleic acid sequences (preferably DNA) and methods for increasing the mRNA levels and protein expression of malarial surface protein MSP-1 which is known to be difficult to express in cell culture systems, mammalian cell culture systems, or in transgenic animals. The preferred protein candidates for expression using the recombinant techniques of the invention are MSP-1 proteins expressed from DNA coding sequences comprising reduced overall AT content or AT rich regions and/or mRNA instability motifs and/or rare codons relative to the native MSP-1 gene.
-
Citations
22 Claims
-
1. A modified nucleic acid sequence comprising SEQ ID NO 1.
-
2. A nucleic acid sequence comprising a modified nucleic acid sequence encoding a merozoite surface protein 1 (MSP-1) operably linked to a promoter which directs expression in mammary epithelial cells wherein the AT content of the modified nucleic acid sequence has been reduced by replacing codons from a nucleic acid sequence of SEQ ID NO:
- 2 with mammary gland preferred codons encoding the same amino acid as the replaced codon such that the AT content of the modified nucleic acid sequence is 50% or less.
- View Dependent Claims (9, 13)
- 3. A vector comprising a modified merozoite surface protein 1 (MSP-1) nucleic acid according to clam 2.
-
10. A nucleic acid sequence comprising a modified nucleic acid sequence encoding a merozoite surface protein 1 (MSP-1) operably linked to a promoter which directs expression in mammary epithelial cells wherein the AT content of the modified nucleic acid sequence has been reduced by replacing codons from a nucleic acid sequence of SEQ ID NO:
- 2 with mammary gland preferred codons such that the AT content of the modified nucleic acid sequence is 50% or less and, wherein at least one glycosylation site of SEQ ID NO;
10 has been altered such that it is not functional. - View Dependent Claims (11, 12, 14)
- 2 with mammary gland preferred codons such that the AT content of the modified nucleic acid sequence is 50% or less and, wherein at least one glycosylation site of SEQ ID NO;
-
15. A nucleic acid comprising a modified nucleic acid encoding merozoite surface protein 1 (MSP-1) operably linked to a promoter which directs expression in mammary epithelial cells, wherein all mRNA instability motifs present in the wild-type nucleic acid sequence encoding MSP-1 have been eliminated by replacing codons of SEQ ID NO:
- 2 with mammary gland preferred codons encoding the same amino acid as the replaced codon.
- View Dependent Claims (16, 17, 18)
-
19. A nucleic acid comprising a modified nucleic acid encoding merozoite surface protein 1 (MSP-1) operably linked to a promoter which directs expression in mammary epithelial cells, wherein all mRNA instability motifs present in the wild-type nucleic acid sequence encoding MSP-1 have been eliminated by replacing codons of SEQ ID NO:
- 2 with mammary gland preferred codons, and wherein at least one glycosylation site of SEQ ID NO;
10 has been altered such that it is not functional. - View Dependent Claims (20, 21, 22)
- 2 with mammary gland preferred codons, and wherein at least one glycosylation site of SEQ ID NO;
Specification