Studies On Gene Clone And Functional Identify As Well As Gene-transfer Of Linalool Synthase Of Lavandula Officinalis

A 600bp sequence was cloned from lavandula officinalis through RT-PCR by using degenerate primers. After comparing with the sequences in Genebank, the sequence was found to share the highest similarity (76%) with the sequence of linalool synthase. The whole cDNA sequence was cloned through 3'RACE and 5'RACE, and the size is 1809bp. The protein sequence contains 603 residues. comparing the protein sequence with the members of monoterpene synthase family, and the result suggested that the sequence contains all of the conserved functional amino acid sequences (DDXXD) which are shared by monoterpene synthases and 5'RR functional groups. The analysis results of nucleic acid and protein sequences suggested that the gene cloned is one member of linalool synthase family, and we name it Llis.getting the full-length cDNA of Llis by using full-length primers. The genome sequence of lavandula officinalis was cloned by using full-length primers, and the size of the sequence is 2320bp. The sequence was found to contain 6 exons and 7 introns. The result of Southern Blotting suggested that in lavandula officinalis there may be two copies of Llis. It was constructed that the prokaryotic expression vector, sense and antisense expression vectors for Llis.The size of the protein produced through prokaryotic expression is neary 60KD, which is equal to the experiment have expected. The expression vectors were transferred into the W38 tobacco, and through ppt selection, extracted genome DNA also PCR amplified. Finally, the positive transgenic tobaccos were got. The result of Northern Blotting suggested that both sense and antisense transgenic tobaccos can express the relevant proteins, and some failed might because of the gene silence.In order to researching on transgenic L ilium spp., it was also used that Lilium spp.as materials to find out the proper conditions for callus induction and differentiation of L ilium spp..