Prokaryote Expression And Preliminary Functional Identification Of DNA Methyltransferases From Rice

DNA methylation is an important epigenetic modification. Cytosine bases of the nuclear genome in higher plants are often extensively methylated. Experimental evidence indicates that cytosine methylation plays important roles in regulating gene expression and maintainting genome integrity.Patterns of cytosine methylation in plants are usually faithfully maintained across generations.It was found that disturbance of intrinsic DNA methylation patterns may lead to functional and phenotypic abnormality, or evolutionary opportunity. On the other hand the inheritance of specific methylation pattern in animals and plants,and variation of methylation induced by biotic or abiotic factors may provide novel insight into the biological significance of this epigenetic marker. Understanding the mechanisms that regulate de novo methylation will give insights into the biological role of genomic methylation patterns, and for this reason it will be necessary to identify and characterize all DNA methyltransferases expressed.In this study, we attempted to clone some fragments which encode de novo DNA methyltransferase into pET-28a vector, induce their expression in E.coli BL21 with IPTG De novo DNA methyltransferase activities were tested with purified recombinant proteins. The results showed that, to this end, only the expression product of DMT710-3 in NEBuffer 1 showed detectable DNA methyltransferase activity.