Clonine Of Methyl-binding Protein MBD701, Construction Of Its Plant Expression Vectors And Genetic Transformation In Rice

DNA Methylation plays important regulation roles in gene expression,cellular differentiation and phylogenetic development. MBD is a kind of trans-actingfactor which binding with DNA methylation, and play a considerable function druingthe growth and development in plant. This research cloned the MBD701in rice andthen constructed on the basis of its RNAi expression vector with different promoters.Some transgentic plantlets were confirmed by preliminary identification.The mainresults are as follows:1. By using RT-PCR cloned cDNA fragment of MBD701of rice, sequences analisisshowed that the full-length1353bp include945bp CDS and5’-UTR19bp as well as3’-UTR389bp, the protein-coding is315amino acids and predicted molecular weightis32.5kD.2. Cluster analysis showed that, MBD701and corn MBD108, MBD111, ArabidopsisAtMBD2, AtMBD12and wheat TaMBD2have the highest homology, which belongto the MBD subgroup III. InterPro analysis of MBD701sequences of genes deducedamino acid indicated that, MBD701contains a typical methyl-binding proteinconserved domain (138~212amino acids) and a CW-type zinc finger domain (73~132amino acids).3. Separately constructed the MBD701RNAi expression vector with35S and PGRpromoter. Used rice as a receptor material, through Agrobacterium-mediated methodto transit constructed RNAi vector for genetic transformation. PCR analysises havebeen performed on the part of the genetic transformation of rice plants, the resultsshowed that: the expected gene integrated into the rice genome have been detected, atotal of58positive plants obtained, which provide a dasis for futher research.