title: Generation and analysis of a large-scale expressed sequence tags from a full-length enriched cDNA library of Siberian tiger (Panthera tigris altaica).
abstract: 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 1.28×10(6)pfu/mL and 1.59×10(10)pfu/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.
authors.affiliation: Department of Bioscience, Department of Laboratory Medicine, Bengbu Medical College, Bengbu 233030, China. Electronic address: ily0720@126.com.
authors.forename: Yu
authors.initials: Y
authors.lastname: Guo
authors.affiliation: Department of Bioscience, Department of Laboratory Medicine, Bengbu Medical College, Bengbu 233030, China; Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China. Electronic address: lcq7813@hotmail.com.
authors.forename: Changqing
authors.initials: C
authors.lastname: Liu
authors.affiliation: Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China. Electronic address: taofenglu@yahoo.com.cn.
authors.forename: Taofeng
authors.initials: T
authors.lastname: Lu
authors.affiliation: The Northeast Tiger Wooden Land of Heilongjiang, Harbin 150028, China. Electronic address: liudan_1964@sina.com.
authors.forename: Dan
authors.initials: D
authors.lastname: Liu
authors.affiliation: Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China; College of Wildlife Resource, Northeast Forestry University, Harbin 150028, China. Electronic address: baichunyu001@hotmail.com.
authors.forename: Chunyu
authors.initials: C
authors.lastname: Bai
authors.affiliation: Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China. Electronic address: nobelli@126.com.
authors.forename: Xiangchen
authors.initials: X
authors.lastname: Li
authors.affiliation: Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China. Electronic address: Yuehui.ma@263.net.
authors.forename: Yuehui
authors.initials: Y
authors.lastname: Ma
authors.affiliation: Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China. Electronic address: wjguan86@hotmail.com.
authors.forename: Weijun
authors.initials: W
authors.lastname: Guan
mesh_terms: D000818:Animals; D002460:Cell Line; D024142:Collagen Type VI / Q000737:chemistry / Q000235:genetics; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D056727:Endangered Species; D020224:Expressed Sequence Tags; D005260:Female; D005347:Fibroblasts; D015870:Gene Expression; D015723:Gene Library; D059785:Karyotype; D008297:Male; D058977:Molecular Sequence Annotation; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics; D017422:Sequence Analysis, DNA; D046010:Tigers / Q000235:genetics*
chemical_list: D024142:Collagen Type VI; D018076:DNA, Complementary; D011993:Recombinant Fusion Proteins
keywords: COL6A2 gene; Expressed sequence tags; Fibroblast cell; SMART cDNA library; Siberian tiger