Identification of cytochrome b Bene of Chinese herb Testudinis Carapax et Plastrum / 中国药学杂志

ABSTRACT

OBJECTIVE: To establish a practical, convenient and accurate method of DNA molecular marker for the identification of Testudinis Carapax et Plastrum. METHODS: Phenol extraction method and salting-out method were used to extract mtDNA from Testudinis Carapax et Plastrum and its adulterants. Cyt b mtDNA sequences of Chinemys reevesii and other turtle species were downloaded from GenBank. Species specific PCR primers were designed according to the differential DNA fragments of Cyt b genes, and Cyt b gene segment was amplified by PCR technique and sequenced. RESULTS: The complete 16.6 kb mtDNA was extracted from all samples by the two extraction methods. The specific primers could only amplify the Cyt b gene sequence of Chinemys reevesii, and its fragment was about 319 bp. The result of sequencing indicated that the homologous similarity was 100% with Chinemys reevesii. CONCLUSION: Salting-out method is suitable for the DNA extraction from Testudinis Carapax et Plastrum. The primers are specific to Chinemys reevesii and can be used to distinguish Testudinis Carapax et Plastrum from its adulterants. Copyright 2012 by the Chinese Pharmaceutical Association.