Construction and evaluation of DNAMethyltransferase vector for RNA interference.

DNA Methylation,the most common form of covalent modification in eukaryotic cell,influencing the interaction between proteins and DNA,plays an important role in the regulation of gene expression.As a new technology for knocking out specific genes,RNAi (RNA interference)is applied extensively to plant functional genomics,development,the regulation of physiological metabolic pathway and so on.in this paper,RNAi expression vectorof DNA methyltransferases (DNMTs) designed based on the conservative sequence DNMTs were constructed,which lays a foundation forthe DNA Methylation regulation of characterimprovement.firstly,the conservative fragment of DNMTs was cloned from the genomic DNA of Arabidopsis.And then a target sequence,about 570bp,was amplified from the received conservative sequence and then the target sequence was inserted into the pHANNIBAL vector.Later the complete ORF contained the inverted repeat was linked to the plant expression vectorPgreenII.By the analysis of both restriction enzyme digestion and sequencing,the accomplishment of the DNMTs-RNAi recombinant expression vectorwas confirmed.The results of further transformation of Taxus cell showed that the RNAi recombinant expression vector had biology function laying a foundation for genetic modification of DNA methylation controlled character.