Hamsters Of The High Variability Of Microsatellite Loci Screening Applications

In this study a partial common great long-tailed hamster(Criecetulus triton) genomic library including more than 4000 clones was constructed. About 74 recombinant positive clones containing microsatellite sequences of C. triton were obtained through rapid screening small size fractionated genomic libraries with PCR technique. The primer sets used in PCR were M13 Primers M3/RV located at both sides of multiple cloning site of pUC19 and (CA)₈, (CT)₈, (AT)₈. Sequencing of these clones was performed with automated sequencer, and 25 microsatellites were isolated. The sequence numbers submitted to the GenBank are AY780293-AY780309, AY864067-AY864074. Among them twenty-two microsatellites have (CA)_n motif, aacounted for 88%, only three microsatellites have (CT)_n motif, accounted for 12%. In the 25 microsatellites, perfect ones are 40%, imperfect 4%, compound perfect 56%. The motif number from 8 to 20 in most microsatelltes are 48%, few motif numbers exceed 25.Ten good microsatellite primer sets were designed based on unique sequences flanking each motif with the software Oligo 6.0 and Primer 5.0. PCR amplification of template DNA of C. triton populations in Shunyi, Gu'an, and Raoyang were carried out with these primers. The result indicated that six microsatellte markers were polymorphism. Cluster analysis showed that there were certain differences bewteen the poulation of Raorang and the populations of shunyi, gu'an. The same microsatellite markers showed different allele number in different population. The dominated alleles and the private alleles found in each population can be different. Primers of PAY7802, PAY7807 and P5022801 can be used to evaluate the genetic structure among different populations as a population character.