细叶石斛的位点特异性PCR鉴别(英文)  

Allele-specific diagnostic PCR authentication of D. hancockii from the other Dendrobium species

作　　者：丁小余[1] 张卫明[2] 保曙琳[1] 常俊[1] 

机构地区：[1]南京师范大学生命科学学院资源生物学重点实验室,南京210097 [2]南京野生植物综合利用研究院,南京210042

出　　处：《中国医学生物技术应用》2002年第4期36-43,共8页The Chinese Academic Medical Magazine of Organisms

摘　　要：根据细叶石斛及其它37种枫斗类和黄草类石斛的rDNAITS序列,我们设计了位点特异性PCR鉴别引物XY-JB01S和XY-JB01X,对细叶石斛进行了成功的DNA分子鉴别。在进行位点特异性鉴别PCR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℃,只有细叶石斛的模板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,已在鉴别细叶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 Dendrobium hancockii and the other 37 species of Dendrobium, thenew allele-specific diagnostic primers XY-JB01S and XY-JB01X have been designed to authenticate D. hancockiifrom the other species. Before the diagnostic PCR, the primers for amplifying the whole ITS region should be used tovalidate template DNA at first so as 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. hancockii could be amplifiedwhereas the diagnostic PCR of the other 37 species were all negative. The diagnostic PCR showed good reproducibil-ity and played an important role in identifying D. hancockii in China. Thus, the allele-specific diagnostic primers havebeen designed to authenticate D. hancockii efficiently. Compared with the authentication method by sequencing DNAfragments, the allele-specific diagnostic PCR was not only simple and timesaving but also practical and effective.

关 键 词：细叶石斛 位点特异性PCR 鉴别 

分 类 号：R284[医药卫生—中药学]