宋聚先,邹佳宁,王晓丽
SONG Ju-xian~1, ZOU Jia-ning~1, WANG Xiao-li~2 (1.Lab for Molecular Pharmacognosy

• 1:贵阳医学院分子生药学实验室
• 2:贵阳医学院分子生药学实验室
• 3:贵阳医学院药用植物学与生药学教研室 贵州贵阳550004
• 4:贵州贵阳550004

摘要(Abstract)：

目的:探讨兰科药用植物天麻(GastrodiaelataBl.)简单重复序列(simplesequencerepeat,SSR)PCR循环中退火温度及反应体系中各主要成分对反应的影响。方法:根据Genebank登录的天麻微卫星序列设计引物,并对反应体系中各影响因素进行优化。结果:所设计引物可扩增出预期的条带,并构建了清晰稳定的SSR指纹图谱,最终建立的25μl反应体系为:TaqDNA酶1U、Mg2+1·2~1·6mmol/L、引物各0·4~0·6μmol/L、模板DNA40~60mg/L和dNTPs0·20mmol/L。结论:首次初步确立了适合天麻SSR的最佳退火温度及反应体系中各主要成分的最佳浓度,为天麻的SSR分子标记及遗传多态性分析奠定了一定基础。
Objective: To observe the effects of annealing temperature of cycling condition and concentration of each element on simple sequence repeat (SSR) reaction system employed for studys of medical plant Gastrodia elata Bl. Methods: Primers were designed according to microsatellite loci of G. elata Bl. recorded in GENEBANK and concentrations of all ingredients of the reaction were optimized. Results: Clear and repeatable SSR fingerprinting is constructed using self-designed primers. The optimized reaction system was: Taq DNA polymarase 1U, Mg~ 2+ 1.2-1.6 mmol.L~ -1 , each primer 0.4-0.6μmol.L~ -1 , template DNA 40-60ng/ul, dNTPs 0.20 mmol.L~ -1 . Conclusions: Optimized annealing temperature and ingredients of SSR reaction system were constructed for the first time which can be used in the molecular fingerprinting and genetic polymorphism analysis.

关键词(KeyWords)： 天麻;序列分析,DNA;串联重复序列
gastrodia, elata;sequence analysis,DNA;tandem repeat sequences

Abstract:

Keywords:

基金项目(Foundation): 科技部2004BA721A33项目;; 贵州省科技黔科合农社字(2001)1125号项目;; 黔科合中药专字[2003]51号项目

作者(Author): 宋聚先,邹佳宁,王晓丽
SONG Ju-xian~1, ZOU Jia-ning~1, WANG Xiao-li~2 (1.Lab for Molecular Pharmacognosy

DOI: 10.19367/j.cnki.1000-2707.2006.06.006

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