| As a kind of important laboratory animal, cats have unique advantage in pharmacology andmedical experiments,especially in neurobiology, cardiovascular and pharmacodynamics.Cats areusually used in observing the reaction in neurological, physiology, pharmacology and toxicologybecause of their experimental results are more closer to humans than rodents. Cats are alsoexcellent materials for cranial nerve physiology for their sensitive nervous system. Furthermorecats are best laboratory animals using in drug screening because their sensitivity to drug is accordwith humans and their blood pressure is more constant than rats and rabbits.Cats’blood vesselwalls are tough enough to be operated so they can be used in analysing the mechanism of drugaction in the circulatory system.There is not uniform standard of genetic monitoring for cats in wordwide though cats areused widely. so the genetic quality could not be well controlled. The laboratory data about cats isnot reliable and accurate considering above.A kind of trustworthy method of genetic monitoringfor cats is needed to established as soon as possible.The objective of our study is to establish themethod of microsatellite genetic monitoring.We obtained131cats microsatellite loci and3sumatran tiger microsatellite loci fromGenbank.Search for the information of primers in related literatures and synthesis theprimers.Firstly,134pairs of primers were performed PCR to the genome DNA of32cats andthen performed1.5%agarose gel electrophoresis to confirm.The principle of screening is to keepthe bandings reasonable and single. Meantime keep a record of the better annealingtemperature.All134microsatellite loci among which there were131cats microsatellite loci and3sumatran tiger microsatellite loci were performed PCR and then Agarose gel electrophoresis.113clear bindings were obtained,in which there were2of all3sumatran tiger microsatellite loci. Thenscreened the polymorphic microsatellite loci. The microsatellite loci that were screened aboveamplificated for the DNA of32domestic cats. A total of31polymorphic microsatellite loci wereobtained.The2sumatran tiger microsatellite loci did not show polymorphic.Synthesis the primerswith fluorescence markers of the31polymorphic microsatellite loci.34laboratory Orange TabbyCats were from North China Pharmaceutical Group New Drug Research and DevelopmentLimited Company.There are no genetic relationships and sexual ignorance in them.The cats were anesthetized by Zoletil then2ml bioclean anticoagulation whole blood samples were drew fromthe vein of Fore leg and extracted the genome DNA from the blood. Next the markedfluorescence primers amplified for34Orange Tabby Cats，genome DNA and then perfomed STRscanning. After obtaining the results of STR scanning we put the genotype of34Orange TabbyCats into popgene3.2to analysis. We got the Mean of effective Allele number, Mean ofShannon’Information index, Mean of effective heterozygosity of polymorphic microsatellite lociof the34Orange Tabby Cats.Afterwards we performed independent sample T-test between theresults from all polymorphic microsatellite loci and the microsatellite loci decreased step by steptill found the lowest numbers of microsatellite. When23microsatellite loci were picked, there wasno significant difference between the Mean of effective Allele number and Mean ofShannon’Information index compared with31microsatellite loci,P>0.05,but the significantdifference was found between the mean of effective heterozygosity compared with31microsatellite loci,P<0.05.So the24microsatellite loci were ascertained to establish the method ofgenetic monitoring for cats.Put the information of genotypes of microsatellite markers that weredetermined to reserve finnaly into Popgene3.2and get the datas of the Average observed numberof alleles, Mean of effective Allele number, Mean of Shannon’Information index, and Mean ofeffective heterozygosity. In our study Average observed number of alleles is4.0833; the mean ofeffective allele number is2.6323; The Mean of Shannon’Information index is1.0786; The mean ofeffective heterozygosity is0.6108which meet the characteristics of a outbred stock (0.4-0.7). The resultsshow that the group of Orange Tabby Cats from North China Pharmaceutical Group New Drug Researchand Development Limited Company accord with the characteristics of a outbred stock. |
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