The Microsatellite Locus Isolation And The Primary Genetic Characterization Analyse Of Blue Fox (Alopex Lagopus)

Blue fox (Alopex lagopus) and silver fox (Vulpes logpus) are for important for fur farming, creating lots of economy benefits every year in China. However, there are many problems in pedigree, inbreeding and descendiblity management. Microsatellite DNAs is a widely used and powerful method in many fields, including population genetic diversity analysis, genomic mapping, QTL allocation, disease diagnosis, research of pathology, paternity testing and individualization. Therefore, the application of the microsatellite method may be necessary in solving the problems of breeding blue fox and silver. However, there are few reports about the microsatellite isolation from fox genome.In this study, genomic DNA was extracted from of blue fox, and then digested with Sau3 A I (Mbo I) before isolation by electrophoresis with 1.5% agarose gel. Fragments of 300bp～900bp was collected, purified and hybridized with (CA)₁₅ probe labeled by Biotin. A microsatellite enriched library was then constructed by using magnetic bead (MagneSphere^? Magnetic Separation Products) and PCR amplifications. The primers were designed according to the map of PMD-18 vector, and PCR reaction was performed with (CA)₁₅N and (TG) ₁₅N for the selection of positive clones and further sequencing. The data of sequencing analysis revealed that 55 microsatellite loci were obtained, and from which 6 loci was chosen to analyz the genetic diversity of 30 blue fox and 30 silver fox. The results showed that in 30 blue fox the value of observed heterozygosity (Ho) among 6 loci ranged from 0.1613 to 0.8125, and the averaging value was 0.4751. The expected value of Ho among 6 loci ranged from 0.245 to 0.8261, and the averaging value was 0.507. Moreover, the effective number of alleles (Ne) of 6 loci ranged from 1.3245 to 5.7515, while the average number was 3.2594. The value of polymorphism information content (PIC) among 6 loci ranged from 0.24 to 0.8001, with the averaging value of 0.5115. In 30 silver fox, the Ne number among 6 loci ranged from 1 to 5.3248, with the averaging value of 2.9578. The observed Ho value ranged from 0 to 0.6, and the averaging value was 0.3404. The expected he value among 6 loci ranged from 0 to 0.8122, with the averaging value of 0.5487. Finally, the PIC value of 6 loci ranged from 0 to 0.7455, with the averaging value of 0.4161.From these results, we conclude that the 6 loci of microsatellite can be used in analysis of the genetic diversity of blue fox and silver fox. Because the genetic diversity of blue fox is better than silver fox, the seed source of silver fox should be improved.