Studying microbial diversity in the frozen soil is crucial to screen and utilize the cold-adapted microorganisms. In this paper, selected the Tianshan permafrost in the unique geographical environment in Xinjiang of China as study material, then diversity of the frozen soil microorganism were determined by pure culture and sequencing. The results indicated that, among 68 isolated strains, 65.2% of them belong to Gram-positive bacteria, and the colony colors were mainly creamy white, lemon yellow, white. The optimum growth temperature of the isolated microorganisms were 20-24°C, the majority of them are cold-adaptable bacteria. after the 16S rDNA sequence analysis, the selected representative strains were subjected to the phylogenetic analyses, these strains cultivable at low temperature belong to 6 phylogenetic groups of a-proteobacteria subclass, b-proteobacteria subclass, g-proteobacteria subclass, Gram-positive bacteria, positive bacteria and CFB, among them, a-, b-, g-proteobacteria was the dominant flora. Pseudomonas spp. is the dominant genus, which accounts for the highest percentage in the grains. We can conclude that permafrost of the Tianshan Mountain can nurture rich diversity of microorganisms at low temperature, and may be viewed as a good source for screening low-temperature microorganism.