Termination of transgene expression via transposon-mediated break
AssigneeNational Taiwan University
InventorYuh-Chyang Charng
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Abstract
This invention provides a genetic construct comprising: (a) a DNA cut-and-paste transposon genetic unit which comprises a transposase gene flanked on either side by inverted repeat sequences and operably under the control of a first promoter; and (b) a transgene unit which comprises an expressable transgene, placed operably under the control of a second promoter; wherein the transposon genetic unit and the transgene unit are linked with one of the inverted repeat sequence located in an intron of the transgene. This invention also provides methods of transiently expressing a transgene in a stably transformed eukaryote. This invention further provides methods for obtaining marker-free transgenic plants.
Claims14
A genetic construct comprising:
- a DNA cut-and-paste transposon genetic unit which comprises a first promoter which is inducible in plants, a transposase gene, a 5′ inverted repeat sequence, which is located in an intron of the transposase gene, a poly(A) fragment, a transgene unit, and a 3′ inverted repeat sequence which are operably linked in the 5′ to 3′ direction, wherein the transposase gene is operably under the control of the first promoter;
- and
- wherein the transgene unit comprises a second promoter and an expressable transgene which are operably linked in the 5′ to 3′ direction, wherein the transgene is operably under the control of the second promoter.
A method of transiently expressing a transgene in a stably transformed plant, comprising the following steps:
- (a) stably transforming the plant with a genetic construct, comprising
- a DNA cut-and-paste transposon genetic unit which comprises a first inducible promoter, a transposase gene, an 5′ inverted repeat sequence, which is located in an intron of the transposase gene, a poly(A) fragement, a transgene unit, and a 3′ inverted repeat sequence which are operably linked in the 5′ to 3′ direction, wherein the transposase gene is operably under the control of the first promoter;
- and
- wherein the transgene unit comprises a second promoter and an expressable transgene which are operably linked in the 5′ to 3′ direction, wherein the transgene is operably under the control of the second promoter;
- (b) expressing the transgene of the genetic construct in the plant;
- and
- (c) expressing the transposase gene of the genetic construct in the plant.
Dependencies are inferred from the claim text.