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为了建立梅花鹿、马鹿及其杂交鹿茸的特异性PCR鉴别方法。采集不同来源的梅花鹿、马鹿鹿及其杂交鹿的鹿角或鹿茸样品共48份,其中24份为市场流通的待测样品。分别利用COI与SRY基因序列作为其父母本的鉴别标记,对已知样品的COI与SRY基因进行扩增、测序,进行同源比对后根据其变异位点设计特异鉴别引物,分别基于COI与SRY基因建立对梅花鹿、马鹿及其杂交鹿的鹿茸的特异PCR鉴定方法,并随机对市场流通的24份待测鹿茸样品进行鉴别分析。本实验所构建双位点特异PCR体系,基于COI的鉴别位点,可通过PCR扩增获得232 bp的梅花鹿特异片段,而产生518 bp的马鹿特异片段;而基于SRY的鉴别位点,通过PCR扩增获得803 bp的梅花鹿特异片段,而产生425 bp的马鹿特异片段。随机抽取市场流通中的24个待测鹿茸样品,经鉴定有3个样品属于马·花杂交的鹿茸样品。本实验建立对梅花鹿、马鹿及其杂交鹿的鹿茸的PCR鉴别方法简便、可靠。
In order to establish a specific PCR identification method for sika deer, red deer and their hybrid antler. A total of 48 antler or antler samples collected from different sources of sika deer, red deer and their hybrid deer were collected, of which 24 were market-tested samples. COI and SRY gene sequences were used as the identification markers of their parents respectively. COI and SRY genes of known samples were amplified and sequenced. After homologous comparison, specific identification primers were designed according to their variation sites. The SRY gene was used to identify the velvet antlers of sika deer, red deer and their hybrid deer, and 24 random samples of pilose antler were tested. In this experiment, a double-site specific PCR system was constructed. Based on the COI-discriminating sites, a 232 bp sika deer specific fragment was obtained by PCR amplification and a 518 bp red deer-specific fragment was obtained. Based on the SRY- PCR amplification of 803 bp sika deer specific fragment, resulting in 425 bp of red deer specific fragment. Twenty-four samples of pilose antler were randomly selected from the market, and three samples were identified as antler samples from hybridized rapeseed. This experiment established the deer Velvet PCR identification of sika deer, red deer and their hybrid deer is simple and reliable.