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

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 p fu/ml and 1.62×109 p fu/ml respectively. The proportion of recombinants from un amplified 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 reveal ed t hat 53. 9% of the sequences were classified as strong match, 38.6% as nominal and 7.4% as weak match. 28. 0 % of t he m were found t o be related t o 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. © Ivyspring International Publisher.