ISSN: 1449-2288International Journal of Biological Sciences
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Int J Biol Sci 2010; 6(6):584-589. doi:10.7150/ijbs.6.584 This issue Cite
Short Research Communication
Construction of cDNA library and preliminary analysis of expressed sequence tags from Siberian tiger
1. Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China
2. Department of Bioscience, Bengbu Medical College, Bengbu 233000, China
3. The Northeast Tiger Wooden Land of Heilongjiang, Harbin 150028, China
* These authors contributed equally to this work.
Abstract
In this study we successfully constructed a full-length cDNA library from Siberian tiger, Panthera tigris altaica, the most well-known wild Animal. Total RNA was extracted from cultured Siberian tiger fibroblasts in vitro. The titers of primary and amplified libraries were 1.30×106 pfu/ml and 1.62×109 pfu/ml respectively. The proportion of recombinants from unamplified library was 90.5% and average length of exogenous inserts was 1.13 kb. A total of 282 individual ESTs with sizes ranging from 328 to 1,142bps were then analyzed the BLASTX score revealed that 53.9% of the sequences were classified as strong match, 38.6% as nominal and 7.4% as weak match. 28.0% of them were found to be related to enzyme/catalytic protein, 20.9% ESTs to metabolism, 13.1% ESTs to transport, 12.1% ESTs to signal transducer/cell communication, 9.9% ESTs to structure protein, 3.9% ESTs to immunity protein/defense metabolism, 3.2% ESTs to cell cycle, and 8.9 ESTs classified as novel genes. These results demonstrated that the reliability and representativeness of the cDNA library attained to the requirements of a standard cDNA library. This library provided a useful platform for the functional genomic research of Siberian tigers.
Keywords: Siberian tiger, Panthera tigris altaica, Fibroblast cell line, SMART cDNA library, Expressed sequence tags.
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