Construction of RNA Interference Vector of TT19 Gene Family in Brassica

The RNAi fragment of Brassica TT19 gene family is cloned by PCR first,and then its sense and antisense fragments are respectively sub-cloned into between promoter and spacer,and between spacer and terminator in platform vector pFGC5941M for forming RNAi vector and finally the RNAi vector is transformed into Agrobacterium tumefaciens strain LBA4404 after PCR identification to lay a foundation for research and modification of testa pigment and ornamental color characters.The sequence results showed that the RNAi fragment(BTT19I) with 200 bp of Brassica TT19 gene family was cloned,which corresponds to 363~540 bp of BnTT19-1 mRNA,its sense and antisense fragments were respectively sub-cloned into pFGC5941M by NcoI+AatII and BamHI+XbaII double digestion and pFGC5941M-BTT19I,a RNAi vector with 10 552 bp was obtained.