ABSTRACT
OBJECTIVE： To establish the method for PCR identification of bullwhip， and to identify the authenticity of bullwhip at the molecular level. METHODS： DNA samples of bullwhip and its counterfeits （donkey whip， pig whip， sheep whip） were extracted and their integrity， purity and concentration were detected. Using GenBank related information， using mitochondrial cytochrome b （Cyt b） gene of bullwhip as target gene， Primer-BLAST online software was used to design specific primer. PCR amplification was performed for whips of different species， and electrophoretic analysis was conducted for the product. PCR products of bullwhip samples were cloned and confirmed by DNA sequencing. The specificity and repeatability of the established PCR method were verified. RESULTS： DNA purity of the bullwhip and its counterfeits was high， and there was no protein or RNA pollution. 1.5％ agarose gel electrophoresis showed that there were obvious target gene bands of bullwhip samples at 200-300 bp， while no corresponding bands appeared in other counterfeit products. The results of DNA sequencing showed that the nucleotide sequence of the gene fragment of bullwhip was 100％ similar to that of the bullwhip in GeneBank. Results of methodological validation showed that established method was specific and reproducible. CONCLUSIONS： The established PCR identification method based on Cyt b gene in the study is simple， rapid， accurate， specific and reproducible， and can meet the requirements of analysis and identification of bullwhip and its counterfeits.
ABSTRACT
We studied the genetic characteristics of Spermophilus in Shaanxi Province,China.The COI,Cyt-b gene were sequenced and the results were compared with those of dauricus from Inner Mongolia Keyouzhong Banner and Zhengxiangbai Banner,and S.alaschanicus from Haiyuan County of Ningxia.And genetic distance was analyzed and Neighbor-Joining tree was built.Results showed that the genetic distance of COI gene sequences between Spermophilus from Dingbian County in Shaanxi and S.alaschanicus in Ningxia was ≤0.5％,and the genetic distance was ranged from 7.9％ to 9.3％ with Citellus dauricus from Inner Mongolia.The genetic distance of Cyt-b gene between Spermophilus from Dingbian County in Shaanxi and S.alaschanicus in Ningxia was ≤2.2％,and ranged from 8.9％ to 11.2％ with Citellus dauricus from Inner Mongolia.The Neighbor-Joining tree of COI,Cyt-b gene showed two major clusters.One of them were clustered by Spermophilus from Dingbian County in Shaanxi and S.alaschanicus in Ningxia,and another one was Citellus dauricus from Inner Mongolia.The Neighbor-Joining tree of COI gene showed that all samples from Shaanxi Province clustered in a group.In conclusion,the Spermophilus in Shaanxi Province were S.alaschanicus.
ABSTRACT
Objective To analyze the Cyt b gene sequences in Tibet mini-pigs and clarify the differences and genetic relationship with other Chinese pigs.Method The sequence of Cyt b gene was amplified from genome DNA of Tibet mini-pig,Bama miniature pigs,Guizhou xiang pigs and Wuzhishan (WZS) pigs.After sequencing,the base sequences were compared and analysed.The blood relationship tree and evolution position of Tibet mini-pig were established.Result There were 14 mutation sites between domestic pigs in China and pigs from Europe.Besides there was a significant differenee in two nucleotide site:a T→C switch in site 420 and the G→A switch in site 883 at the same time.Conclusion Chinese pigs include Bama miniature pigs,Guizhou xiang pigs and WZS pigs,have a very close blood relationship with some of Tibet mini-pigs.It has been confirmed that there is a certain genetic differentiation in the Tibet mini-pig.
ABSTRACT
Objective To differentiate the sequences of Cyt b gene Agkistrodon from its adulterants sold in the market in order to provide molecular evidence for identification of Agkistrodon. Methods Cyt b gene was used to sequence and analyze Agkistrodon and its adulterants sold in the market. Results There was a mixed phenomenon in Agkistrodon species. The differences of Cyt b gene sequence between Agkistrodon and its adulterants sold in the market are significant: the difference rates among the species of Agkistrodon are 0%—0.91%, the difference rates between Agkistrodon and its adulterants are 18.57%—23.78%. Conclusion The characteristics of Cyt b gene sequence can be used as a better molecular marker for authenticating Agkistrodon from its adulterants.