Identification of bear bile powder and its adulterants by using DNA barcoding technology / 中国中药杂志

To identify the precious bile powder and its adulterants by DNA barcoding, and establish its standard experimental process to ensure the safe and effective utilization. Total twelve sequences from samples of bear bile powder which come from Ursus thibetanus for DNA extraction, PCR(polymerase chain reaction) and sequence, then using CodonCode Aligner V 7.0.1 shear primer region to obtain COI sequence. The COI sequences of U. arctos and their adulterants were obtained from GenBank. MEGA7.0 software was applied for analyzing mutation, calculating intraspecific and interspecific K2P(Kimura 2-Parameter) genetic distance and constructing the Neighbor-joining tree(NJ). The results showed that the maximum K2P genetic distance of bear bile powder of U. thibetanus and U. arctos are far less than minimum K2P genetic distance within its adulterants species, and the results of NJ tree demonstrated that each species could be distinguished from the counterfeits obviously. DNA barcoding is a safe, convenient and reliable technique for species identification, and it is important to establish the standard sequence of COI sequences for animal medicines.

Identification of Medicinal Insect Vespa mandarinia from Its Adulterants Using DNA Barcode / 世界科学技术-中医药现代化

This study aimed at exploring a fast method to accurately identify the medicinal insect Vespa mandarinia Smith from its adulterants using DNA barcode and COI sequences.The extracted DNAs from V.mandarinia and its adulterants V.soror were amplified by polymerase chain reaction (PCR) and sequenced bilaterally based on COI barcode sequence investigation.The information of the COI sequences of V.mandarinia and V.soror were gathered from GenBank.All the sequences were compared and analyzed,and their intraspecific and interspecific genetic distances were calculated using MEGA 6.06.In addition,the phylogenetic tree was established with neighbor-joining (NJ) method.As a result,the COI sequences of V.mandarinia and V.soror were successfully amplified.The minimum interspecific distance between V.mandarinia and its adulterants was 0.152 ± 0.017,being considerably larger than the maximal intraspecific distance between V.mandarinia,0.009±0.004.The constructed phylogenetic tree showed an independent branch for each species.It was concluded that the DNA barcode based on COI sequence can efficiently identify V.mandarinia and its adulterants.This study provided an innovative tool for the quality control and market regulation of Chinese materia medica,securing the safe medication of V.mandarinia.

Molecular identification of hippocampus and its adulterants based on COI barcode / 中国药学杂志

OBJECTIVE: To establish an accurate identification method of genuine Hippocampus based on COI sequences as DNA barcodes and analyze the relationship between the genuine and adulterants of Hippocampus. METHODS: Two hundred and e-leven pieces of COI sequences were collected from 21 Hippocampus species in this study. After PCR amplification and sequencing of the DNAs, the DNA barcoding gap and phylogenetic cluster analysis were carried out. RESULTS: The COI sequences of the five authentic species all had obvious DNA barcoding gap. For phylogenetic cluster analysis, all of the samples were monophyletic and every species could be discriminated clearly. CONCLUSION: COI is an effective DNA barcode for the identification of the genuine Hippocampus and its adulterants, and have important significance for clinical medication safety.

Identification of the skin of Bufo bufo gargarizans and its adulterants based on DNA barcoding / 中国药学杂志

OBJECTIVE: To provide new technique for quick and accurate identification of the skin of Bufo bufo gargarizans u-sing DNA barcoding method based on the COI sequence. METHODS: Fifty-four samples of the skin of Bufo bufo gargarizans and its four kinds of adulterants were collected from 16 different localities. The DNAs were extracted, the COI squences were amplified and sequenced bi-directionally. The obtained sequences were assembled using the CodonCode Aligner V 4.2. Then the genetic distances were accumulated using Kimura-2-Parameter(K2P) model, and the NJ tree was constructed by MEGA5.0. RESULTS: The intra-specific genetic distances were much smaller than inter-specific ones between the skin of Bufo bufo gargarizans and its adulterants. Furthermore, the NJ tree showed that the skin of Bufo bufo gargarizans can be distinguished from its adulterants clearly. CONCLUSION: The COI sequence can be used as the DNA barcode to identify the skin of Bufo bufo gargarizans and its adulterants with accuracy and efficiency.