The Black Line Hamster (cricetulus Barabensis) Microsatellite Primers Were Screened And Applications

Partial small - insert genomic library composed of more than 3000 clones was screened with PCR technique for recombinant positive clones containing microsatellite sequences of Cricetulus barabensis . The primers used in PCR were M13 Primers M3/M13 Primers RV which located at both sides of multiple clone site of pUC19 and microsatellite primers (CA)₈/(GA)₈designed according to core repeats motifs of microsatellite . 65 recombinant positive clones containing microsatellite obtained by screening were sequenced and 60 microsatellite loci were found from 38 clones. GenBank accession numbers of these clones are DQ235662, DQ270160, DQ270163, DQ270166, DQ270170-DQ270172 , DQ286857-DQ286860, DQ270173-DQ270177, DQ342058-DQ342061, DQ459486-DQ459503. Among these microsatellite loci there are 36 perfect ones in 60%;6 imperfect ones in 10%;18 compound ones in 30%.Repeat number of most microsatellites varies from 4 to 20,few microsatellites exceed 20.Nine microsatellite sequences were selected for designing oligonucleotide primers, using Lasergene 6 and Primer 5.0 software .After extensive optimization, 5 pairs of primers gave satisfactory results and were polymorphism. Genetic polymorphism of Cricetulus barabensis populations in Qufu Wucun ,Qufu Fangshan tumen and Pingyi Mengshan were studied with these primers. The result indicated that genetic variation of Pingyi Mengshan population were highest.Cluster analysis showed that there were differences between the Pingyi Mengshan population and Qufu Wucun ,Qufu Fangshan tumen population ,but differences between three populations were small.