| The red panda (Ailurus fulgens) is one of the critically endangered animals today in the worldwide conservation and it was listed in the Appendix I of the CITES (Convention on International Trade in Endangered Species of Wild Flora and Fauna). Due to poaching for red panda and the disturbance of habitat, it's populations are redued sharply, and are becoming seriously endangerous. The global population of the red pandas is estimated only about 16, 000-20, 000 (Choudhury 2001; Wei et al. 1999). Now there are many researches in morphologic, taxology, histology and ecology of red panda, However, little research about molecular biology of it, especially in genetic analysis.Microsatellite DNA is co-dominate genetic marker, it can fealty reflect personal feature, the parentage relationship, it have been wildly used in human and livestock. In this time, more microsatellites loci in red panda were found and the characterizes of these loci also analyzed, The genetic diversity of the three captive red panda populations were analyzed also.In this research we used the method of Bloor et al.(2001) and make some changes. We chose three oligonucleotide (CAA)8, (GGA)8 and (GATA)6. According to the strong affinity between streptavidin and biotin, this three oligonucleotide labeled with biotin, respectively. These three probes were then reacted with streptavidin-coated magnetic beads. The genomic DNA of red panda were digested with restriction enzyme Sau3AI. The 300-1000bp DNA fragments were isolated, ligated with linkers Oligo A and Oligo B, then annealed with streptavidin-biotin-probe complexes. PCR amplificated the streptavidin-riched DNA, the flagments with 300-1000bp were reclaimed and ligated with pGEM-T easy vector, After transformated the ligated DNA to Escherichia coli JM109, induced by X-Gal and IPTG, white bacterium were picked out. Then we constructed 3 enriched libraries (CAA)n, (GGA)n and (GATA)n. Each of the library have a few thousand of clones, Positive clones were screened with PCR amplification from the(CAA)n, (GGA)n and (GATA)n libraries. The results showed that the libraries have 4%, 4.4%, 7.8% positive clones, respectively. From sequence we found these loci: (CAA)n three, (GGA)n eleven, (GATA)n thirty-seven, (TCC)n one, (CCA)n two, (AC) twenty-seven。And 28 loci all have high polymorphisms and good for future analyzed. So, they were used to analysis the three populations of Panda base, Pi county, and Kunming zoo.The number of alleles per locus ranged from 2 to 15, with anaverage of 9.09 in Panda base, 6.69 in Pi xian and 3.72 in Kunming zoo. The observed and expected heterozygosities ranged from 0.050～0.950 and 0.251～0.899, from 0.111～1 and 0.442～0.953, from 0～1and 0～1 repectively. The polymorphic information content (PIC) values for these loci was from 0.195 (Aifu-5, Kunming zoo) to 0.897 (Aifu-31, Pixian). Acorrding to Botstein, when the PIC was abve 0.5, the locus should be a high polymorphic marker; And the PIC was between 0.25 to 0.5, the locus should be a middle polymorphic marker. So in the 28 loci, Aifu-18, Aifu-23, Aifu-29, Aifu-35 belongs to middle polymorphic markers, others belongs to high polymorphisms, and are good DNA maekers for future analyzed.
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