Construction of expression vector of ethylene receptor genefrom banana for RNA interfering

A 538 bp DNA fragments downstream the AUG start codon of the banana ethylene receptor gene was amplified by PCR in both sense and anti-sense orientation. A vector that contained the inverted-repeat DNA fragment was constructed. After sequencing demonstration,the inverted-repeat DNA fragment was cloned into pCAMBIA-1301 vector in which it replaced the GUS gene under control of the 35S promoter. In addition,some questions,such as the linking difficulty between target inserting fragment and cloning vector resulted from intra-strand annealing during constructing vectors including inverted repeat complementary sequence,were discussed.