中文摘要：

黄芩是一种传统的中药,黄芩苷是其重要的药用成分之一,具有广泛的药理作用并被用于多种疾病的治疗,具有巨大的药用价值和经济价值。CHS是催化黄芩苷合成限速步骤的关键酶,其表达量对黄芩苷的积累起关键的作用。本研究利用基因工程的手段,构建黄芩的CHS过表达载体,并将其导入根癌农杆菌中,拟在黄芩中持续表达CHS,促进黄芩苷合成。基于NCBI上公布的黄芩基因信息,设计特异引物,通过RT-PCR技术扩增CHS的c DNA全长序列,并利用生物信息学软件对序列进行分析,在引物两端引入特异酶切位点,将目的序列CHS通过限制性内切酶酶切和连接的方式,构建载体p BI 121-35S-CHS。通过PCR和酶切的方法进行验证,获得黄芩CHS基因的高效表达载体,可直接用于黄芩的遗传转化,使CHS基因表达量增加,促进黄芩苷的合成。

英文摘要：

Scutellaria baicalensis Georgi is a traditional Chinese medicine, as one of the medicinal components, baicalin has the great medicinal value and economic value since it shows wide pharmacologic activities and has been applied extensively in disease cure. Chalcone synthase （CHS） is the key enzyme catalyzing the rate-limiting step of baicalin biosynthesis, its expression level directly affects the accumulation of baicalin. In this study, CHS overexpression vector was constructed and introduced into A grobacterium tumefaciens, the objectives of this study was to improve the biosynthesis of baicalin by overexpression of CHS gene. The CHS sequence was download from NCBI, CHS full length eDNA sequence was obtained by RT-PCR using specific primers. Sequence analysis of CHS was finished by bioinformatics software, then restriction enzyme cutting site was added into the end of each primer and CHS sequence was introduced into the pBI121-35S vector. After PCR and restriction enzyme digestion verification, the pBI121-35S-CHS overexpression vector was constructed successfully and could be used in Scutellaria baicalensis Georgi genetic transformation to increase the amount of CHS gene expression and improve the accumulation ofbaicalin.