Sponsoring Institution
Agricultural Research Service/USDA
Status
ACTIVE
Funding Source
USDA INHOUSE
Reporting Frequency
Annual
Project Director
STOVER E W
Accession Number
425662
Project Number
6034-21000-014-22T
Dates
2013-10-01 - 2016-05-01
Recipient Organization
AGRICULTURAL RESEARCH SERVICE
219 SOUTH ROCK ROAD
FT PIERCE,FL 34945
Keywords
biotechnology
citrus
clean
diseases
gene
hlb
recombination
site-specific
technology
trangenic
transformation
Research Effort
Applied (50%)
Basic (50%)
Developmental (0%)
Classification Parameters
| Knowledge Area | Subject of Investigation | Field of Science | Percent |
|---|
| 204 - Plant Product Quality and Utility (Preharvest) |
920 - Orange |
1080 - Genetics (excludes breeding) |
40% |
| 203 - Plant Biological Efficiency and Abiotic Stresses Affecting Plants |
910 - Grapefruit |
1000 - Biochemistry and biophysics |
30% |
| 212 - Pathogens and Nematodes Affecting Plants |
999 - Citrus, general/other |
1160 - Pathology |
30% |
Goals / Objectives
The research objectives are to reduce potentially negative effects of transgene insertion and genomic presence. The aim of this proposed research is to investigate the use of novel unidirectional recombinases Bxbl, CinH. ParA and phiC31 to implement a Recombinase-Mediate Cassette Exchange (RMCE) technique for precise integration with simultaneous marker removal.
- To test the newly generated transgenic founder citrus Carrizo citrange lines containing the RMCE genetic platform for precise targeting.
- To demonstrate that the dual unidirectional RMCE can be exchanged for constructs with desirable transgenes in Carrizo citrange.
Methods (unparsed)
Transgenic Carrizo citrange founder lines have been generated containing the selection gene cassette flanked by fused recognition sites. Single copy-number, highly expressing lines will be selected. An exchange vector will be transformed into the various Carrizo citrange founder lines. Recombinase mediated cassette exchange will be examined by negative selection and used to score the most effective pairs. The most effective Carrizo citrange founder lines and recombinase pairs will be published and made publicly available.
Methods
Transgenic Carrizo citrange founder lines have been generated containing the selection gene cassette flanked by fused recognition sites. Single copy-number, highly expressing lines will be selected. An exchange vector will be transformed into the various Carrizo citrange founder lines. Recombinase mediated cassette exchange will be examined by negative selection and used to score the most effective pairs. The most effective Carrizo citrange founder lines and recombinase pairs will be published and made publicly available.