Molecular Cloning And Expression Profiling Of A Chalcone Synthase Gene And Phenylalanie Ammonia-lyase From Lamiophlomis Rotate

In this study, a phenylalanine ammonia-lyase(PAL) gene and a chalcone synthase(CHS) gene was cloned from Lamiophlomis rotate of YUSHU by the methods of Reverse-Transcription PCR and RACE,And bioinformatics analysis of the gene sequence.Using the method of Real- Time PCR detection the expression characteristics of PAL gene and CHS gene in organizations of Lamiophlomis rotate.In order to reveal the regulation function of these two genes in different organs,molecular theory foundation to the research of resistance and pharmacy.the main results of this paper studies were as follows: 1、The full-length cDNA of Lamiophlomis rotate PAL designated as LrPAL(GenBank No. KF385881)was 2298 bp,and contained a complete open reading frame(ORF) of 2145 bp, which encoded 714 amino acid residues. The full-length genomic DNA of LrCHS(Gen Bank No. KM114220) was 2026 bp with one intron of 651 bp and two exons of 178 bp and 998 bp. The cDNA sequence of LrCHS gene had an 1176 bp open reading frame encoding a 391 amino acid protein.Sequence analysis showed than it contains the typical PAL enzyme active site sequence. 2、Homology analysis indicated that the deduced LrPAL protein was highly homologoud to other PAL proteins from different species. CHS protein predicted from L. rotate showed 79%-86% identities with CHS of other plant species and retained most of the conserved residues.Phylogenetic tree analysis revealed that LrPAL had closer relationship with PALs from Labiatae plants than from other plants.,and LrCHS was located in Labiatae branch. 3、The expression level of CHS gene in different organization of L. rotate showed flowers> leaves > petioles > roots > stems trend, and the expression level of PAL gene showed petioles > leaves > flowers > roots > stems trend. This assay could successfully provide a technical platform to research L. rotate genes at the mRNA levels in the quantitative analysis.These results alone blindly resistance and molecular foundation for pharmaceutical research.