The Effective Ingredient Synthetase Gene Cloning Of Coleus Forskohlii( Wild. ) And The Ecological Environm Ent Study

Qiaoruisu is the dry grass of Coleus forskohlii(wild.). The major production place of this crude medicine is tropical region or subtropics like India, Sri Lanka and Nepal. It is also found in china, Yunnan. In nongovernmental we used it to cure asthma and tumour,and it show great curative effect, so we called it almighty medicine.On account of the enormous medicinal value. Hubei Furen Pharmacy Incorporated Company creat new traditional Chinese medicine by Qiaoruisu, common hogfennei root and liquorice, and introduce a fine variety of Coleus forskohlii( wild.) from Yunnan to Hubei Tongcheng.The previous research results of our laboratory room had show that Thongcheng cultivate Coleus forskohlii(wild.)has the same pharmacologic action with Yunnan Coleus forskohlii(wild.),but the content of the main index sign ingredients diterpenoid compounds isoforskolin is lower than Yunnan Coleus forskohlii(wild.).This paper had studied Qiaoruisu effective ingredient synthetase gene and the dependency of the content and entironment. In order to improve Hubei Tongcheng plant Qiaoruisu active ingredient concentration.1.Using Tongcheng Qiaoruisu planting base grow Qiaoruisu blade as testing piece. Building Qiaoruisu Kauri acid oxidase template and using the result of Sanger sequencing to design primer. Using PCR technology to get whole of the Qiaoruisu blade Kauri acid oxidase gene. The result show that the total of Qiaoruisu Kauri acid oxidase gene is 1500 bp, and the number of amino acid is 500, belong to CYP450 subfamily, the amino acid sequence has some homology of other species.2. Using the Kauri acid oxidase gene sequence to design primer and add restriction enzyme cutting site Hind Ⅲ, XbaⅠ. Through PCR technology amplification the gene and shift to pYES2 yeast strain, build expression vector and do bioinformatics analysis. The result show that, the Kauri acid oxidase gene has high similarity with other plant,it has 78% similarity with Til plants seasame seeds, has 80% similarity with Scrophulariaceae plants wait crispy noodles, has 73% similarity with Salvia miltiorriza bunge, has 71% similarity with Rubiaceae coffee plants, has 87.11% similarity with Chinese chestuut fafaceae plants, the isoelectric point is 9.10, the aliphatic index is 87.11, the molecular formula is C2566H4015N707O712S24. Kauri acid oxidase is hyfrophilic protein and distribution in external lamina. 3. TO compare the environment of Yunnan Coleus forskohlii and Hubei Coleus forskohlii Planting Base. Investigate the effects of forskolin Coleus growth Soil and climatic factors, In order to improve the Tongcheng planting herbs Coleus active ingredient content. Using modern analytical methods to measured two place soil pH value, Organic matter, Nitrogen content and Phosphorus content. Through literature review and on-site investigation, comprehensive analysis of the differences with the City of Tongcheng and Yunnan Luquan ecological environment,combined Coleus forskohlii active ingredient content differences to discuss the correlation of soil nutrient and active ingredient content. Results: The soil of Yunnan is lack of nutrient than tongcheng and pH value is more acid. So the result of this study had provide reference for Hubei Tongcheng standardized planting Qiaoruisu.