What is the restriction enzyme for Ti plasmid?
More than two RE’s are used for Ti plasmid. Technique of Transferring Plant Gene Using Ti Plasmid – The simplest way to introduce T -DNA into plant cells is to infect them with A. tumefaciens containing the appropriate Ti plasmid and let nature do the rest. Therefore, we need to be able to insert desired genes into the T-regions of Ti plasmids. Since the Ti plasmid is very large (up to 235 kbp), it is not feasible to modify it directly; hence it is useful to perform all manipulations on an excised piece of DNA including the T-DNA and then use in vivo recombination to swap the engineered T-DNA for its normal version in an intact Ti plasmid. The strategy developed is as follows. First, the T-region is cut out of a Ti plasmid with restriction enzymes and introduced into one of the standard cloning vector plasmids that are used with E. coli. Large amounts of the vector carrying the T -DNA can be grown in E. coli and then isolated. The next step is to use restriction enzymes and recombinant
