珊瑚菜Gl4CL 基因克隆与生物信息学分析

目的：对动物药材DNA提取方法进行优化，利用优化方法提取市售动物药材DNA并进行DNA条形码鉴定。方法：基于SDS法DNA提取原理，比较裂解液中不同EDTA浓度（0.025、0.25、0.5 mol·L-1）、是否含NaCl和Triton X-100等因素对不同用药部位动物药材DNA提取质量的影响，筛选得到最佳裂解液配方；使用优化的裂解液配方提取121份市售动物药材DNA并进行基原物种鉴定。结果：裂解液配方为1 % SDS、0.03 mol·L-1 Tris-HCl、0.25 mol·L-1 EDTA、0.2 mol·L-1NaCl对不同用药部位动物药材DNA提取效果最佳，并可实现对蝉蜕等提取困难样本DNA的提取；利用优化裂解液提取的121份市售动物药材DNA满足中药材分子鉴定后续实验要求，所有市售动物药材均可准确鉴定到基原物种。结论：本研究优化的裂解液配方可用于除壳类、分泌物类、加工品外不同用药部位动物药材的DNA提取，为动物药材分子鉴定提供了技术支持。

Cloning and Bioinformatics Analysis of Gl4CL Gene in Glehnia littoralis

The DNA extraction method of animal medicine material is difficult and un- unified, which limits the application of molecular identification to identify animal medicines. In this study, based on the DNA extraction theory of SDS, we assessed the effects of three elements including different EDTA concentrations (0.025 mol·L-1, 0.25 mol·L-1,and 0.5 mol·L-1) and whether containing NaCl and Triton X-100 in the lysis buffer on the quality of DNA extracted fromdifferent kinds of animal medicine. The optimized lysis buffer was used to extract DNA from 121 commercial animal medicines for original and species identification. The results showed that the lysis buffer of 1% SDS, 0.03 mol·L-1 Tris-HCl, 0.25 mol·L-1 EDTA and 0.2 mol·L-1 NaCl had the optimum effect on DNA extraction. This lysis buffer can obtain DNA from animal medicine which is difficult to extract, such as Cicadae periostracum. The DNA extractions of 121 commercial animal medicines by optimized lysis buffer can satisfy the experimental requirements for molecular identification. All samples of commercial animal medicines can be accurately identified to the level of species. It was concluded that optimized lysis buffer can be used in the DNA extraction of different kinds of animal medicines except shells, secretions and processed products. This method provides technique support for the molecular identification of animal medicines.