Construction of cDNA library and preliminary analysis of expressed sequence tags from Siberian tiger

文献类型： 外文期刊

第一作者： Liu, Chang-Qing

作者： Liu, Chang-Qing;Lu, Tao-Feng;Feng, Bao-Gang;Guan, Wei-Jun;Ma, Yue-Hui;Liu, Chang-Qing;Liu, Dan

作者机构：

关键词： Siberian tiger;Panthera tigris altaica;Fibroblast cell line;SMART cDNA library;Expressed sequence tags

期刊名称：INTERNATIONAL JOURNAL OF BIOLOGICAL SCIENCES （ 影响因子：6.58； 五年影响因子：6.478 ）

ISSN： 1449-2288

年卷期： 2010 年 6 卷 6 期

页码：

收录情况： SCI

摘要： 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 1.30x10(6) pfu/ml and 1.62x10(9) 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 the m 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.

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