The Correlation Between HQT Gene Polymorphism And The Content Of Chlorogenic Acid Of Lonicera Japonica And Near Source&HQT Gene Eukaryotic Expression Vector Construction

The flower buds of Lonicera japonica Thunb. are widely used in Traditional Chinese Medicine for the treatment of wet heat illness, the common cold, sore throat, pneumonia, erysipelas, the honeycomb tissue inflammation, dysentery and other diseases, honeysuckle has been widely used because its efficacy and significant effect.However, some lower efficacy, morphologically similar honeysuckle closely related species also as a medicinal plant used in the clinical treatment.In order to the10th edition of Pharmacopoeia, Lonicera japonica Thunb. as the only source of the Lonicera japonica plant, and Lonicera including Lonicera macranthoides Hand.-Mazz., Lonicera confusa DC., Lonicera hypoglauca Miq., and Lonicera fulvotomentosa P.S.Hsu..Morphological similarity, marketing and practical application, often act as honeysuckle Lonicera confusa or mistakenly used as honeysuckle Lonicera confusa phenomenon, which will inevitably lead to the efficacy deviation.The objective of this research is to reveal the HQT gene polymorphism of Lonicera japonica and near souurce, and elaborate the correlation between HQT gene polymorphism and the content of chlorogenic acid. Some varieties of Lonicera plants were used as materials that came from different origin and near source, the HQT gene coding region was amplified by RT-PCR; sequencing; the multiple compare was used to analyse the HQT gene polymorphism; the HPLC was used to determine the sample’s content of chlorogenic acid; then the correlation between HQT gene polymorphism and the content of chlorogenic acid was analysed. There were two kinds of mutation in these HQT gene coding sequences, including base substitution mutations, insertion/deletion mutation; and there was no mutant in conserved region of HQT amino acid sequence. The polymorphism of HQT gene was quite abundant in Lonicera japonica and near source, and there was certain degree of correlation between HQT gene polymorphism and the content of chlorogenic acid. On the based of HQT gene’s coding sequence, used gateway cloning system completed the built of HQT gene eukaryotic expression system, while the completion of the construction and optimization of genetically modified tissue culture system.Therefore, this study systematically honeysuckle and its near-source species HQT gene variants with chlorogenic acid content and HQT gene expression in eukaryotes a preliminary study, to fill a honeysuckle HQT gene bioinformatics research gaps, scientific the use and development of honeysuckle resources provides an important theoretical support and technical means.