Simultaneous excision of two transgene flanking sequences and resolution of complex integration loci |
| |
Authors: | Sandeep Kumar William F Thompson |
| |
Institution: | Plant Gene Expression Lab, Plant Biology Department, NC State University, Campus Box 7550, Raleigh, NC 27695, USA. skumar3@unity.ncsu.edu |
| |
Abstract: | In planta excision techniques have proven useful both for basic biology and applied biotechnology. In this report, we describe a simple site-specific recombination (SSR) strategy that simultaneously removes pre-defined DNA sequences from both sides of a transgenic "gene of interest," leaving only the desired gene and short sequences from the recombinase recognition site. We have used the FLP/FRT SSR system to provide a proof of concept, though any of several other SSR systems could be used in the same way. The frequency of double excision ranged from 33% to 83% in different transgenic lines. We show that a single SSR reaction can simultaneously carry out double excisions and resolve complex transgene loci at high frequency. The method has direct biotechnological application and provides a useful tool for basic research. |
| |
Keywords: | |
本文献已被 PubMed SpringerLink 等数据库收录! |
|