兜唇石斛的位点特异性PCR鉴别  

Allele-specific Diagnostic PCR Authentication of D.aphyllum from the Other Dendrobium Species

作　　者：丁小余[1] 徐珞珊[2] 常俊[1] 保曙琳[1] 丁秉中[1] 

机构地区：[1]江苏省资源生物技术重点实验室 [2]中国药科大学中药学院生药学研究室,南京210038

出　　处：《南京师大学报（自然科学版）》2002年第4期71-76,共6页Journal of Nanjing Normal University(Natural Science Edition)

基　　金：江苏省资源生物技术重点实验室开放基金;国家自然科学基金(30171144)资助课题.

摘　　要：　根据兜唇石斛及其它37种枫斗类和黄草类石斛的rDNAITS序列,设计了鉴别兜唇石斛的位点特异性PCR引物DC JB01S和DC JB01X,并对兜唇石斛成功地进行了位点特异性PCR鉴别.在进行位点特异性PCR鉴别之前,首先运用扩增ITS区的通用引物P1、P2对模板DNA进行扩增,以验证模板的可靠性和扩增的合适浓度.当退火温度上升为64℃,只有兜唇石斛的模板DNA能被扩增出来,而其它的37种石斛属植物均为阴性.该鉴别反应重复性好,已在鉴别我国兜唇石斛中发挥重要作用.与DNA测序鉴别方法相比,位点特异性PCR具有简单、省时、高效、准确等优点.Based on rDNA ITS sequences of D.aphyllum and the other 37 species of Dendrobium,the new allelespecific diagnostic primers DCJB01S and DCJB01X have been designed to authenticate D.aphyllum from the other species.Before the diagnostic PCR,the primers for amplifying the whole ITS region should be used to validate template DNA first to obtain the appropriate template DNA concentration for the diagnostic PCR.When the annealing temperature was raised to 64℃,only the template DNA of D.aphyllum could be amplified whereas the diagnostic PCR of the other 37 species were all negative.The diagnostic PCR have been repeated for many times and have played an important role in identifying D.aphyllum in China.Compared with the authentication method by sequencing DNA fragments,the allelespecific diagnostic PCR was not only simple and timesaving but also effective and accurate.

关 键 词：兜唇石斛 位点特异性PCR 鉴别 药材 

分 类 号：R282.71[医药卫生—中药学] R282[医药卫生—中医学]