Construction Of Rice OsTOM1-RNAi Expression Constructs And Trangenic Rice Plants Expressing OsTOM1-RNAi

The full-length cDNA and genomic DNA sequence of Rice OsTOM1gene which was from TP309, a breed of rice, has been cloned by themethods of RACE and specific primers based on the rice EST sequenceAU173304(gi:12623091) of rice which was searched by blasting thesequence of AtTOM1 of Arabidopsis thaliana in the GeneBank. The lengthof the genomic sequence of OsTOM1 gene is 7140bp long with 11 exons and10 introns. The full-length cDNA of Rice OsTOM1 gene is 1341bp long witha ORF(Open Reading Frame)from the 156th base pair to the 1062th basepair, encoding a 302 amino acids putative protein. there are 155 base pairsat the 5'non-coding region and 279 base pairs at the 3'non-coding regionrespectively. Antisense orientation and RNAi expression constructs weredesigned based on the OsTOM1 gene. The transgenes were introduced intorice varity TP309 by Agrobacterium tumefaciens- mediated transformation.Twelve transgenic plants that were transformed with the RNAi constructand resistance to G418 were obtainted and The results of PCRdemonstrated that the target genes were integrated into the genome of riceplants. The whole cDNA sequence and complete genomic DNA sequence ofthe gene pave the way for further studying the function of the OsTOM1 gene. And the antisense orientation and RNAi expression constructs lay afoundation for getting the anti-virus transgene rice.