关键词: Cyt b基因, DNA序列, PCR鉴定, 蛇, 药材

Abstract: In order to develop a more applicable DNA molecular marker identification method for the authentication of Bungarus parvus and its adulterants, DNA templates were extracted from Bungarus parvus, adulterants of the crude drugs and their original. animals. Cyt b gene fragment was amplified from all these templates respectively using universal primers. PCR products were purified and sequenced directly. After sequence alignment and comparing the sequence of Bungarus parvus to its adulterants, we find that Cyt b gene is a good molecular marker for authentication of the crude drugs since interspecific differentiation is more evident than that of intraspecies in DNA sequence of the gene. Based on the sequence data of Cyt b gene fragment, a pair of highly specialized primers, used as diagnostic primers, was designed for the PCR identification of the crude drugs. Using diagnostic primers for the PCR identification, Bungarus parvus samples could be absolutely distinguished from all its adulterers when annealing temperature is at 60 ºC~65 ºC, and no incorrect or missing discrimination was found under these PCR conditions. Mixed powder of the qualified crude drug and its adulterants could also be detected by diagnostic PCR. The results indicate that diagnostic PCR using the primers designed in the present study is a simple, effective and applicable method for the authentication of Bungarus parvus, and this method might also be a new way for examining the compositions of Chinese patent medicine.

Key words: Cyt b gene, Cyt b gene, DNA sequence, PCR identification, DNA sequence, PCR identification, Snake, Snake, Crude drugs, Crude drugs