Claims
- 1. A method of determining the successful transformation of a plant with a heterologous gene, comprising:
transforming a first plant with a target DNA construct comprising at least one heterologous gene operatively associated with a morphological marker that encodes a normal phenotypical characteristic of the plant, wherein the first plant is homozygous for a mutation of the morphological marker that renders the plant abnormal for the phenotypical characteristic; collecting seeds from the first plant and growing at least one progeny plant; and then detecting the phenotype of the progeny plant, wherein the presence of the normal phenotype as indicated by the characteristic encoded by the morphological marker indicates the successful transformation of the progeny plant with the heterologous gene.
- 2. The method of claim 1, wherein the normal phenotypical characteristic is the presence of trichomes on the plant, and the first plant does not have trichomes.
- 3. The method of claim 1, wherein the first plant is homozygous for a mutation of the GL1 gene.
- 4. The method of claim 3, wherein the first plant is homozygous for the gl1 mutation.
- 5. The method of claim 3, wherein the heterologous gene of the DNA construct is operatively associated with the GL1 gene.
- 6. The method of claim 1, wherein the first plant is homozygous for a mutation of the Cot1 gene.
- 7. The method of claim 5, wherein the heterologous gene of the DNA construct is operatively associated with the Cot1 gene.
- 8. The method of claim 1, wherein the transformation of the first plant is carried out by a method selected from the group consisting of direct gene transfer into protoplasts, microprojectile bombardment, injection into protoplasts, cultured cells and tissues or meristematic tissues, electroporation, and Agrobacterium-mediated transformation.
- 9. The method of claim 1, wherein the transformation is carried out by Agrobacterium-mediated transformation.
- 10. The method of claim 1, wherein the first plant comprises a driver DNA construct that transactivates the target DNA construct.
- 11. The method of claim 10, wherein the first plant is homozygous for the driver DNA construct.
- 12. The method of claim 10, wherein the transformation occurs by sexually crossing the first plant carrying a driver DNA construct with a second plant comprising the target DNA construct.
- 13. The method according to claim 10, wherein the transformation occurs by transforming the first plant with the target DNA construct.
- 14. The method of claim 10, wherein the driver DNA construct comprises an activation domain and a DNA binding domain, wherein the DNA binding domain recognizes the target DNA construct.
- 15. The method according to claim 14, wherein the driver DNA construct further comprises a strong promoter.
- 16. The method according to claim 1, wherein the target DNA construct is under the control of a minimal promoter.
- 17. The method according to claim 1, wherein the first plant is Arabidopsis thaliana.
- 18. The method according to claim 1, wherein the target DNA construct is carried on an Agrobacterium binary vector.
- 19. The method according to claim 19, wherein the Agrobacterium binary vector is PPG360.
- 20. The method according to claim 10, wherein the driver DNA construct has been transformed into the first plant by Agrobacterium-mediated transformation.
- 21. A method of detecting the successful transformation of a Arabidopsis plant with a heterologous gene, comprising:
transforming a first Arabidopsis plant that is homozygous for a mutation of the GL1 gene with a target DNA construct comprising a heterologous gene operatively associated with the wild-type GL1 gene; allowing the first Arabidopsis plant to produce seeds; growing the seeds into progeny Arabidopsis plants; and detecting the presence or absence of trichomes on the progeny Arabidopsis plant, wherein the presence of trichomes indicates the successful transformation of the progeny plant with the heterologous gene.
- 22. The method of claim 21, wherein the first Arabidopsis plant is homozygous for the gl1-1 mutation.
- 23. The method of claim 21, wherein the first Arabidopsis plant is homozygous for a mutation of the Cot1 gene.
- 24. The method of claim 21, wherein the transformation of the first Arabidopsis plant is carried out by a method selected from the group consisting of. direct gene transfer into protoplasts, microprojectile bombardment, injection into protoplasts, cultured cells and tissues or meristematic tissues, electroporation, and Agrobacterium-mediated transformation.
- 25. The method of claim 21, wherein the transformation is carried out by Agrobacterium-mediated transformation.
- 26. The method of claim 21, wherein the first Arabidopsis plant comprises a driver DNA construct that transactivates the target DNA construct.
- 27. The method of claim 26, wherein the first Arabidopsis plant is homozygous for the driver DNA construct.
- 28. The method of claim 26, wherein the transformation occurs by sexually crossing the first plant carrying a driver DNA construct with a second plant Arabidopsis plant comprising the target DNA construct.
- 29. The method according to claim 26, wherein the transformation occurs by transforming the first plant with the target DNA construct.
- 30. The method of claim 26, wherein the driver DNA construct comprises an activation domain and a DNA binding domain, wherein the DNA binding domain recognizes the target DNA construct.
- 31. The method according to claim 30, wherein the driver DNA construct further comprises a strong promoter.
- 32. The method according to claim 21, wherein the target DNA construct is under the control of a minimal promoter.
- 33. The method according to claim 21, wherein the target DNA construct is carried on an Agrobacterium binary vector.
- 34. The method according to claim 33, wherein the Agrobacterium binary vector is PPG360.
- 35. The method according to claim 26, wherein the driver DNA construct has been transformed into the first plant by Agrobacterium-mediated transformation.
- 36. A plant useful for determining the successful transformation of a heterologous gene, wherein the plant is homozygous for a mutation of a morphological marker that renders the plant abnormal for the phenotypical characteristic, and wherein the plant further comprises a target DNA construct comprising at least one heterologous gene operatively associated with the morphological marker that encodes a normal phenotypical characteristic of the plant.
- 37. A seed produced by the plant of claim 36 being crossed with a plant
- 38. A progeny plant grown from the seed of claim 37.
- 39. The plant of claim 36, wherein the plant is Arabidopsis thaliana.
- 40. The plant of claim 36, wherein the plant is homozygous for the gl1 mutation.
- 41. The plant of claim 21, wherein the first Arabidopsis plant is homozygous for a mutation of the Cot1 gene.
- 42. The plant of claim 36, wherein the morphological marker is the presence of trichomes.
- 43. The plant of claim 36, wherein the plant comprises a driver DNA construct that transactivates the target DNA construct.
- 44. The plant of claim 36, wherein the plant is homozygous for the driver DNA construct.
- 45. A method of determining the successful transformation of a plant with a nucleotide sequence, comprising:
first plant has an abnormal phenotype for a morphological marker; collecting seeds from the first plant and growing at least one progeny plant; and then detecting the phenotype of the progeny plant, wherein the presence of the normal phenotype of the morphological marker of the progeny plant indicates the successful transformation of the progeny plant with the nucleotide sequence.
- 46. The method according to claim 45, wherein the nucleotide sequence comprises DNA.
- 47. The method according to claim 45, wherein the nucleotide sequence is a heterologous gene.
- 48. The method according to claim 45, wherein the morphological marker is the presence of trichomes.
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application claims the benefit of U.S. Provisional Application No. 60/196,753 filed Apr. 12, 2000.
Provisional Applications (1)
|
Number |
Date |
Country |
|
60196753 |
Apr 2000 |
US |