Cloning and differential expression of chalcone synthase gene 1 in Houttuynia cordata and its protein sequence analysis

WEI Lin;WU Xian-jin;LIU Sheng-gui;TANG Yu-lian;HE An-na;XU Jie;WU Cheng-jiu;Key Laboratory of Xiangxi Medicinal Plant and Ethnobotany of Hunan Higher Education,Key Laboratory of Research and Utilization of Ethnomedicinal Plant Resources of Hunan Province,Department of Life Sciences,Huaihua University;College of Agronomy,Sichuan Agricultural University;Xiangya Medical Laboratory,Central South University;  

Objective To clone and sequence the chalcone synthase1(CHS1) gene from the leaves of Houttuynia cordata.Methods The cloning primers were designed on the basis of the conserved sequences of the cloned CHS gene in other plants.The total RNA was extracted from the leaves of H.cordata.The sequence of CHS gene was cloned by reverse transcription polymerase chain reaction(RT-PCR) and SON-PCR techiniques,and then the gene was ligated with pMD18-T Simple vector.The positive clone was sequenced after the identification of clone by PCR.Results We cloned a fragment of 1 188 bp.The analysis of sequencing indicated that the fragment encoded 395 amino acids and shared the sequence homology of more than 62.3% with CHS gene sequences from other higher plants.Bioinformatic analysis indicated that the CHS amino acids had no signal peptide sequences,but possessed the CHS family's characteristic sequences,RLMMYQQGCFAGGTVLR and GVLFGFGPGL.Relative real-time PCR analysis indicated that CHS1 showed the highest transcript abundance in the flowers,moderate levels in the stems,lower levels in the rhizomes,and the lowest levels in the leaves.Conclusion It is the first report that a novel CHS gene is cloned from H.cordata,which lays a foundation for the effective use of CHS1 gene.

【Fund】： 国家自然科学基金资助项目(30870230);; 湖南省高校创新平台开放基金项目(12K132);; 湖南省科技计划重点项目(2013FJ6090)
【CateGory Index】： R29

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