Recombinant promoter with increased fiber-specific expression
The present application discloses a recombinant fiber-selective promoter region comprising a DNA molecule comprising a fiber specificity region of a cotton lipid transfer protein gene promoter, operably linked to a DNA molecule comprising a nucleotide sequence having at least 90% sequence identity to a nucleotide sequence of about 500 consecutive nucleotides of the 3′ end of the FB8-like 2 promoter and use thereof to increase fiber-selective expression of products of interest in cotton fiber cells.
1. A recombinant DNA molecule comprising a chimeric promoter, which comprises in order:
(i) a DNA molecule comprising a fiber specificity region of a cotton lipid transfer protein gene promoter comprising a nucleotide sequence having the nucleotide sequence of SEQ ID NO: 4, operably linked to
(ii) a DNA molecule comprising a nucleotide sequence having about 500 consecutive nucleotides of the 3′end of the FB8-like 2 promoter of SEQ ID NO: 2, and wherein the chimeric promoter has the cotton fiber specific promoter activity of the synthetic promoter as set forth in SEQ ID NO: 1.
2. The recombinant DNA molecule of claim 1 , wherein the DNA molecule of part (i) comprising the fiber specificity region is preceded by a DNA molecule comprising a nucleotide sequence of about 400 consecutive nucleotides of the 5′end of the FB8-like 2 promoter of SEQ ID NO: 2.
3. The recombinant DNA molecule of claim 1 , wherein the nucleotide sequence of about 500 consecutive nucleotides of the 3′ end of the FB8-like 2 promoter of SEQ ID NO: 2 comprises the nucleotide sequence of SEQ ID NO: 2 from the nucleotide at position 427 to the nucleotide at position 922.
4. The recombinant DNA molecule of claim 1 , wherein the nucleotide sequence of about 500 consecutive nucleotides of the 3′ end of the FB8-like 2 promoter of SEQ ID NO: 2 comprises the nucleotide sequence of SEQ ID NO: 5 from the nucleotide at position 911 to the nucleotide at position 1405.
5. The recombinant DNA molecule of claim 1 , wherein the nucleotide sequence of about 500 consecutive nucleotides of the 3′ end of the FB8-like 2 promoter of SEQ ID NO: 2 comprises the nucleotide sequence of SEQ ID NO: 6 from the nucleotide at position 3638 to the nucleotide at position 4132.
6. The recombinant DNA molecule of claim 1 , wherein the nucleotide sequence of about 500 consecutive nucleotides of the 3′ end of the FB8-like 2 promoter of SEQ ID NO: 2 comprises the nucleotide sequence of SEQ ID NO: 7 from the nucleotide at position 1781 to the nucleotide at position 2276.
7. The recombinant DNA molecule of claim 2 , wherein the DNA molecule comprising a nucleotide sequence of about 400 consecutive nucleotides of the 5′ end of the FB8-like 2 promoter of SEQ ID NO: 2 comprises the nucleotide sequence of SEQ ID NO: 5 from the nucleotide at position 61 to the nucleotide at position 475.
8. The recombinant DNA molecule of claim 2 , wherein the DNA molecule comprising a nucleotide sequence of about 400 consecutive nucleotides of the 5′ end of the FB8-like 2 promoter of SEQ ID NO: 2 comprises the nucleotide sequence of SEQ ID NO: 6 from the nucleotide at position 2787 to the nucleotide at position 3202.
9. The recombinant DNA molecule of claim 2 , wherein the DNA molecule comprising a nucleotide sequence of about 400 consecutive nucleotides of the 5′ end of the FB8-like 2 promoter of SEQ ID NO: 2 comprises the nucleotide sequence of SEQ ID NO: 7 from the nucleotide at position 1047 to the nucleotide at position 1464.
10. The recombinant DNA molecule of claim 1 , wherein the chimeric promoter comprises a nucleotide sequence having at least 98% nucleotide sequence identity to the nucleotide sequence of SEQ ID NO: 1 [from the nucleotide at position 1 to the nucleotide at position 1053].
11. The recombinant DNA molecule of claim 1 , wherein said chimeric promoter has cotton fiber-selective expression of a coding region operably linked thereto.
12. A recombinant nucleic acid construct comprising the following operably linked DNA molecules:
(a) a recombinant DNA molecule according to claim 1 ;
(b) a DNA molecule encoding a biologically active RNA molecule; and
(c) a DNA molecule encoding a transcription termination region or a transcription termination and polyadenylation region.
13. The recombinant nucleic acid construct of claim 12 , wherein said DNA molecule encoding said biologically active RNA molecule codes for a chitin synthase 2 of Neurospora crassa or a glutamine:fructose-6-phosphate amidotransferase of E. coli.
14. A cotton plant cell transformed with a recombinant nucleic acid construct according to claim 12 .
15. A cotton plant transformed with a recombinant nucleic acid construct according to claim 12 .
16. A method for making a transgenic cotton plant cell or cotton plant comprising transforming a cell of a cotton plant with a recombinant nucleic acid construct according to claim 12 , and regenerating a transformed cotton plant from said transformed cotton plant cell.
17. The method of claim 16 comprising further the step of harvesting fibers from said transformed cotton plant.
18. A fiber obtained from the transformed cotton plant of claim 15 , wherein the fiber comprises the recombinant nucleic acid construct.