Claims for Patent: 5,859,347

Summary for Patent: 5,859,347

Title:	Enhanced expression in plants
Abstract:	This invention provides HSP70 introns that when present in a non-translated leader of a chimeric gene enhance expression in plants.
Inventor(s):	Brown; Sherri Marie (Chesterfield, MO), Santino; Colleen Gail (St. Louis, MO)
Assignee:	Monsanto Company (St. Louis, MO)
Application Number:	08/758,149
Patent Claims:	1. An isolated DNA segment comprising in sequence: (a) a promotor that functions in monocotyledonous plant cells; (b) a non-translated leader DNA comprising an intron sequence including at least sufficient nucleotide sequence from the 5' end and from the 3' end of SEQ ID NO: 1 to splice said intron sequence; and (c) a DNA sequence that in combination with (a) and (b) effects production of an RNA sequence; wherein the non-translated leader of (b) increases expression of the DNA sequence relative to wild-type expression. 2. The isolated DNA segment of claim 1, further comprising a 3' non-translated sequence that functions in plant cells to cause addition of a polyadenylated nucleotide sequence to the 3' end of said RNA sequence. 3. The isolated DNA segment of claim 1, wherein said intron sequence comprises at least about 162 nucleotides created by ligating a 5' end sequence and a 3' end sequence from SEQ ID NO:1. 4. The isolated DNA segment of claim 3, wherein said intron sequence is SEQ ID NO:3. 5. The isolated DNA segment of claim 3, wherein said intron sequence comprises at least about 283 nucleotides created by ligating a 5' end sequence and a 3' end sequence from SEQ ID NO: 1. 6. The isolated DNA segment of claim 5, wherein said intron sequence is SEQ ID NO:2. 7. The isolated DNA segment of claim 5, wherein said intron sequence is SEQ ID NO: 1. 8. The isolated DNA segment of claim 1, fused to said leader DNA further comprising between about 10 and about 50 additional nucleotides of an exon fused to said leader DNA. 9. The isolated DNA segment of claim 1, further comprising between about 11 and about 28 additional nucleotides of an exon fused to said leader DNA. 10. The isolated DNA segment of claim 1, further comprising between about 10 and about 50 additional nucleotides from the 3' end of HSP70 exon 1 and between about 11 and about 28 additional nucleotides from the 5' end of HSP70 exon 2 fused to said leader DNA. 11. The isolated DNA segment of claim 1, wherein said intron sequence comprises a splice site consensus sequence. 12. The isolated DNA segment of claim 11, wherein said intron sequence comprises a 5' splice site consensus sequence. 13. The isolated DNA segment of claim 11, wherein said intron sequence comprises a 3' splice site consensus sequence. 14. The isolated DNA segment of claim 11, wherein said intron sequence comprises a 5' splice site consensus sequence and a 3' splice site consensus sequence. 15. The isolated DNA segment of claim 11, wherein said intron sequence has an adenine nucleotide at position 15 of SEQ ID NO: 1. 16. The isolated DNA segment of claim 11, wherein said intron sequence has an adenine nucleotide at position 20 of SEQ ID NO: 1. 17. The isolated DNA segment of claim 11, wherein said intron sequence has a guanine nucleotide at position 800 of SEQ ID NO: 1. 18. The isolated DNA segment of claim 14, wherein said intron sequence has an adenine nucleotide at position 15, an adenine nucleotide at position 20, and a guanine nucleotide at position 800 of SEQ ID NO: 1. 19. The isolated DNA segment of claim 1, wherein said promoter comprises a plant DNA virus promoter. 20. The isolated DNA segment of claim 14, wherein said promoter is a CaMV35S promoter or an FMV promoter. 21. The isolated DNA segment of claim 1, wherein said DNA sequence encodes an EPSP synthase, a CP4 protein, an ACC-deaminase, a B.t. crystal toxin, a glgC16 protein, a plant viral coat protein or a GOX protein. 22. A method for the expression of a gene in a monocotyledonous plant, comprising introducing into the plant cell a DNA segment comprising in sequence: (a) a promotor that functions in monocotyledonous plant cells; (b) a non-translated leader comprising an intron sequence including at least sufficient nucleotide sequence from the 5' end and from the 3' end of SEQ ID NO: 1 to splice said intron; and (c) a DNA sequence comprising said gene; wherein the DNA segment comprising (a), (b) and (c) produces an RNA sequence that effects expression of said gene. 23. The method of claim 22, wherein said DNA further comprises a 3' non-translated sequence that functions in monocotyledonous plant cells to cause addition of a polyadenylated nucleotide sequence to the 3' end of said RNA sequence. 24. The method of claim 22, wherein said intron sequence comprises at least about 162 nucleotides comprised by ligating a 5' end sequence and a 3' end sequence from SEQ ID NO: 1. 25. The method of claim 22, further comprising between about 10 and about 50 additional nucleotides of an exon fused to said leader DNA. 26. The method of claim 22, wherein said intron sequence comprises a splice site consensus sequence. 27. The method of claim 22, wherein said DNA sequence encodes an EPSP synthase, a CP4 protein, an ACC-deaminase, a B.t. crystal toxin, a glgC16 protein, a plant viral coat protein or a GOX protein. 28. The method of claim 22, wherein said plant is maize, wheat or rice. 29. A transgenic monocotyledonous plant comprising the DNA of claim 11. 30. The plant of claim 29, wherein said DNA further comprises a 3' non-translated sequence that functions in plant cells to cause addition of a polyadenylated nucleotide sequence to the 3' end of said RNA sequence. 31. The plant of claim 29, wherein said intron sequence comprises at least about 162 nucleotides obtained by ligating a 5.degree. end sequence and a 3.degree. end sequence from SEQ ID NO: 1. 32. The plant of claim 29, further comprising between about 10 and about 50 additional nucleotides of an exon fused to said leader DNA. 33. The plant of claim 29, wherein said intron sequence comprises a splice site consensus sequence. 34. The plant of claim 29, wherein said DNA sequence encodes an EPSP synthase, a CP4 protein, an ACC-deaminase, a B.t. crystal toxin, a glgC16 protein, a plant viral coat protein or a GOX protein. 35. The plant of claim 29, wherein said plant is maize, wheat or rice.

Details for Patent 5,859,347

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	For Injection	103132	06/04/1986		2007-01-14
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	For Injection	103132			2007-01-14
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	Injection	103132			2007-01-14