IUCN / SSC Cat Specialist Group - Digital Cat Library
   

 

View printer friendly
Liu, C.; Liu, D.; Guo, Y.; Lu, T.; Li, X.; Zhang, M.; Ma, J.; Ma, Y.; Guan, W.
Construction of a full-length enriched cDNA library and preliminary analysis of expressed sequence tags from Bengal tiger
2013  International Journal of Molecular Sciences (14): 11072-11083

In this study, a full-length enriched cDNA library was successfully constructed from Bengal tiger, _Panthera tigris tigris_, the most well-known wild Animal. Total RNA was extracted from cultured Bengal tiger fibroblasts _in vitro_. The titers of primary and amplified libraries were 1.28 ž 106 pfu/mL and 1.56 x 109 pfu/mL respectively. The percentage of recombinants from unamplified library was 90.2% and average length of exogenous inserts was 0.98 kb. A total of 212 individual ESTs with sizes ranging from 356 to 1108 bps were then analyzed. The BLASTX score revealed that 48.1% of the sequences were classified as a strong match, 45.3% as nominal and 6.6% as a weak match. Among the ESTs with known putative function, 26.4% ESTs were found to be related to all kinds of metabolisms, 19.3% ESTs to information storage and processing, 11.3% ESTs to posttranslational modification, protein turnover, chaperones, 11.3% ESTs to transport, 9.9% ESTs to signal transducer/cell communication, 9.0% ESTs to structure protein, 3.8% ESTs to cell cycle, and only 6.6% ESTs classified as novel genes. By EST sequencing, a full-length gene coding ferritin was identified and characterized. The recombinant plasmid pET32a-TAT-Ferritin was constructed, coded for the TAT-Ferritin fusion protein with two 6x His-tags in _N _and _C_-terminal. After BCA assay, the concentration of soluble Trx-TAT-Ferritin recombinant protein was 2.32 ñ 0.12 mg/mL. 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 genome and transcriptome research of Bengal tigers.

PDF files are only accessible to Friends of the Cat Group. Joining Friends of the Cat Group gives you unlimited access and downloads in the Cat SG Library for one year, and allows you to receive our newsletter Cat News (2 regular issues per year plus special issues). More information how to join here

 

(c) IUCN/SSC Cat Specialist Group ( IUCN - The World Conservation Union)