title: Construction and analysis of Siberian tiger bacterial artificial chromosome library with approximately 6.5-fold genome equivalent coverage.
abstract: Bacterial artificial chromosome (BAC) libraries are extremely valuable for the genome-wide genetic dissection of complex organisms. The Siberian tiger, one of the most well-known wild primitive carnivores in China, is an endangered animal. In order to promote research on its genome, a high-redundancy BAC library of the Siberian tiger was constructed and characterized. The library is divided into two sub-libraries prepared from blood cells and two sub-libraries prepared from fibroblasts. This BAC library contains 153,600 individually archived clones; for PCR-based screening of the library, BACs were placed into 40 superpools of 10 × 384-deep well microplates. The average insert size of BAC clones was estimated to be 116.5 kb, representing approximately 6.46 genome equivalents of the haploid genome and affording a 98.86% statistical probability of obtaining at least one clone containing a unique DNA sequence. Screening the library with 19 microsatellite markers and a SRY sequence revealed that each of these markers were present in the library; the average number of positive clones per marker was 6.74 (range 2 to 12), consistent with 6.46 coverage of the tiger genome. Additionally, we identified 72 microsatellite markers that could potentially be used as genetic markers. This BAC library will serve as a valuable resource for physical mapping, comparative genomic study and large-scale genome sequencing in the tiger.
authors.affiliation: Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China. ily0720@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. baichunyu001@hotmail.com.
authors.forename: Chunyu
authors.initials: C
authors.lastname: Bai
authors.affiliation: Department of Bioscience, Bengbu Medical College, Bengbu 233000, China. ily0720@hotmail.com.
authors.forename: Yu
authors.initials: Y
authors.lastname: Guo
authors.affiliation: The Siberian Tiger Park of Heilongjiang, Harbin 150028, China. liudan1964@gmail.com.
authors.forename: Dan
authors.initials: D
authors.lastname: Liu
authors.affiliation: Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China. taofenglu@gmail.com.
authors.forename: Taofeng
authors.initials: T
authors.lastname: Lu
authors.affiliation: Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China. nobelli@gmail.com.
authors.forename: Xiangchen
authors.initials: X
authors.lastname: Li
authors.affiliation: College of Wildlife Resource, Northeast Forestry University, Harbin 150028, China. jianzhangma01@gmail.com.
authors.forename: Jianzhang
authors.initials: J
authors.lastname: Ma
authors.affiliation: Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China. Yuehui_Ma@hotmail.com.
authors.forename: Yuehui
authors.initials: Y
authors.lastname: Ma
authors.affiliation: Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China. wjguan86@hotmail.com.
authors.forename: Weijun
authors.initials: W
authors.lastname: Guan
mesh_terms: D000818:Animals; D002460:Cell Line; D022202:Chromosomes, Artificial, Bacterial / Q000235:genetics*; D056727:Endangered Species; D016678:Genome / Q000235:genetics*; D015698:Genomic Library; D023281:Genomics / Q000379:methods; D018895:Microsatellite Repeats / Q000235:genetics; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA; D046010:Tigers / Q000235:genetics*