Founder Lines for Improved Citrus Biotechnology
Crop Improvement & Utilization Research
Project Number: 5325-21000-020-04
Start Date: Oct 01, 2011
End Date: Sep 30, 2014
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 Bxb1, CinH, ParA and phiC31 to implement a Recombinase-Mediated Cassette Exchange (RMCE) technique for precise integration with simultaneous marker removal.
1) To generate transgenic founder citrus Carrizo citrange lines containing the RMCE genetic platform for precise targeting.
2) To demonstrate proof of concept that the dual unidirectional RMCE is functional in Carrizo citrange.
Single copy transgenic Carrizo citrange founder lines will be generated containing the selection gene cassette flanked by fused recognition sites. An exchange vector will be biolistcally 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.