The Northeast Tiger Wooden Land of Heilongjiang

Harbin, China

The Northeast Tiger Wooden Land of Heilongjiang

Harbin, China
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Guo Y.,Bengbu Medical College | Liu C.Q.,Bengbu Medical College | Xu L.S.,Bengbu Medical College | Lu T.F.,Chinese Academy of Agricultural Sciences | And 5 more authors.
Journal of Animal and Plant Sciences | Year: 2015

Tiger is a special species only found in Asia and considered as one of the world’s most endangered species. In this study, a full-length enriched cDNA library was successfully constructed from Siberian tiger. The titers of primary and amplified libraries were 1.28×106 pfu/ml and 1.59×1010 pfu/ml respectively. The proportion of recombinants from unamplified library was 91.3% and average length of exogenous inserts was 1.06 kb. A total of 365 individual ESTs with sizes ranging from 264 to 1,378bps were then analyzed. Furthermore, 305 unigenes were successfully annotated and involved in 47 functions of the GO classification, Cell (278, 91.1%), Cellular process (302, 82.6%) and Binding (250, 82.0%) are the dominant terms. A total of 234 unigenes were assigned to 244 KEGG pathways, and the pathways with most representation are metabolic pathways (40, 17.1%). Among the 25 COG categories, general function prediction only cluster (52, 14.2%) represents the largest group, followed by translation, ribosomal structure and biogenesis (41, 11.2%), replication, recombination and repair (30, 8.2%), and only 9.9% ESTs (36) classified as novel genes. Moreover, this library provided a useful platform for the functional genome and transcriptome research for the P. tigris and other felid animals in future. © 2014, Pakistan Agricultural Scientists Forum. All rights reserved.


Guo Y.,Bengbu Medical College | Liu C.,Bengbu Medical College | Liu C.,Chinese Academy of Agricultural Sciences | Lu T.,Chinese Academy of Agricultural Sciences | And 6 more authors.
Gene | Year: 2014

In this study, a full-length enriched cDNA library was successfully constructed from Siberian tiger, the world's most endangered species. The titers of primary and amplified libraries were 1.28×106pfu/mL and 1.59×1010pfu/mL respectively. The proportion of recombinants from unamplified library was 91.3% and the average length of exogenous inserts was 1.06kb. A total of 279 individual ESTs with sizes ranging from 316 to 1258bps were then analyzed. Furthermore, 204 unigenes were successfully annotated and involved in 49 functions of the GO classification, cell (175, 85.5%), cellular process (165, 80.9%), and binding (152, 74.5%) are the dominant terms. 198 unigenes were assigned to 156 KEGG pathways, and the pathways with the most representation are metabolic pathways (18, 9.1%). The proportion pattern of each COG subcategory was similar among Panthera tigris altaica, P. tigris tigris and Homo sapiens, and general function prediction only cluster (44, 15.8%) represents the largest group, followed by translation, ribosomal structure and biogenesis (33, 11.8%), replication, recombination and repair (24, 8.6%), and only 7.2% ESTs classified as novel genes. Moreover, the recombinant plasmid pET32a-TAT-COL6A2 was constructed, coded for the Trx-TAT-COL6A2 fusion protein with two 6× His-tags in N and C-terminal. After BCA assay, the concentration of soluble Trx-TAT-COL6A2 recombinant protein was 2.64±0.18mg/mL. This library will provide a useful platform for the functional genome and transcriptome research of for the P. tigris and other felid animals in the future. © 2014 Elsevier B.V.


Liu C.,Chinese Academy of Agricultural Sciences | Liu C.,Bengbu Medical College | Liu D.,The Northeast Tiger Wooden Land of Heilongjiang | Guo Y.,Bengbu Medical College | And 6 more authors.
International Journal of Molecular Sciences | Year: 2013

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 × 106 pfu/mL and 1.56 × 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 6× His-tags in N and C-terminal. After BCA assay, the concentration of soluble Trx-TAT-Ferritin recombinant protein was 2.32 ± 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. © 2013 by the authors; licensee MDPI, Basel, Switzerland.


Liu C.-Q.,Chinese Academy of Agricultural Sciences | Liu C.-Q.,Bengbu Medical College | Lu T.-F.,Chinese Academy of Agricultural Sciences | Feng B.-G.,Chinese Academy of Agricultural Sciences | And 3 more authors.
International Journal of Biological Sciences | Year: 2010

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.

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