中药材乌梢蛇及其混淆品的DNA序列分析鉴别

AUTHENTICATION OF THE CHINESE CRUDE DRUG “WUSHAOSHE” ( ZAOCYS DHUMNADES ) AND ITS SUBSTITUTES BY DNA SEQUENCE ANALYSIS

ＤＮＡ序列分析鉴别是中药材品种鉴定的新方法［１］，已应用于海马、龟甲、鳖甲等中药材的鉴定［２～７］。目前商品流通中乌梢蛇（ＺＡＯＣＹＳ）药材的混淆品种类较多，游蛇科（Ｃｏｌｕｂｒｉｄａｅ）的常见种类都有可能被当作乌梢蛇制成药材，品种鉴定困难［８］。而...

AIM: To distinguish the commercial crude drug “wushaoshe” ( Zaocys dhumnades ) from its substitutes, especially when they were cut into pieces, is difficult. In order to solve the problem, the Cyt b gene fragment both of “Wushaoshe” and of its substitutes together with their corresponding original animals were sequenced in the present research. METHODS: Cyt b gene fragment of about 308 bps was amplified using a pair of universal primers, L14841 and H15149. The PCR products were purified and then sequenced by silver staining DNA sequencing technique. RESULTS: DNA sequence of 246 bps Cyt b gene fragment were obtained from 22 individuals including Zaocys dhumnades and 9 other species of common colubrid snakes. Among the sequence data, 101 varied sites were found. The interspecific substitution varies from 11 84% to 23 98% with an average of 16 85%. The intraspecific substitution varies from 0 to 4 06% with an average of 1 14%. The former is fifteen times of the latter. CONCLUSION: The Cyt b gene fragments were highly conservative at intraspecific level in the snake, while they were less conservative at interspecific level. Hence, the sequence of this fragment is a good molecular marker for identification of the crude snake drugs.