Abstract:

Abstract：With the Gateway technology based on homologous recombination and for the request of pENTR TOPO vector and LCHS2 gene sequence from Oriental Lily cv 'Sorbonne', a special pair of primers were designed, the upstream of which beared 'CACC' at 5' end. An interference fragment of 636bp was obtained by PCR amplification with the primers above and cloned into pENTR/D-TOPO to form pENTR/D-CHS via TOPO cloning. Then the ccdB fragment on destination vector pHellsgate12 was replaced by the interference fragment on pENTR/D-CHS via LR reaction to form RNAi expression vector pH12-CHSi, which was transformed into Agrobacterium strain GV3101 by electroporation. The finished work provides foundation for petunia transformation and research on the effect of down-regulated expression of CHS gene on pigmentation of flowers.