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Báo cáo sinh học: "Construction and characterization of a BAC library from a gynogenetic channel catfish Ictalurus punctatus"

Tuyển tập các báo cáo nghiên cứu về sinh học được đăng trên tạp chí sinh học thế giới đề tài: Construction and characterization of a BAC library from a gynogenetic channel catfish Ictalurus punctatus | 613 Genet Sei. Evol. 35 2003 673-683 INRA EDP Sciences 2003 DOI 10.1051 gse 2003046 Original article Construction and characterization of a BAC library from a gynogenetic channel catfish ỉctaỉurus punctatus SYlvie M.A. Quinioua b TakaYuki Katagiria Norman w. Millera Melanie Wilsona William R. Woltersb GeoffreY c. Waldbieserb a University of Mississippi Medical Center Department of Microbiology Jackson MS 39216 USA b U.S. Department of Agriculture Agricultural Research Service Catfish Genetics Research Unit Thad Cochran National Warmwater Aquaculture Center Stoneville MS 38776 USA Received 4 December 2002 accepted 9 April 2003 Abstract - a bacterial artificial chromosome BAC library was constructed by cloning Hindlll-digested high molecular weight DNA from a gynogenetic channel catfish Ictalurus punctatus into the vector pBeloBACll. Approximately 53 500 clones were arrayed in 384-well plates and stored at 80 C CCBL1 while clones from a smaller insert size fraction were stored at 80 C without arraying CCBL2 . Pulsed-field gel electrophoresis of 100 clones after Noll digestion revealed an average insert size of 165 kb for CCBL1 and 113 kb for CCBL2. Further characterization of CCBL1 demonstrated that 10 of the clones did not contain an insert. CCBL1 provides a 7.2-fold coverage of the channel catfish haploid genome. PCR-based screening demonstrated that 68 out of 74 unique loci were present in the library. This represents a 92 chance to find a unique sequence. These libraries will be useful for physical mapping of the channel catfish genome and identification of genes controlling major traits in this economically important species. bacterial artificial chromosome catfish genome Ictalurus punctatus 1. introduction Bacterial artificial chromosome BAC libraries have become a widespread tool for maintaining entire genomes as large DNA insert clones due to stability low levels of chimeric inserts and ease of manipulation. BAC libraries can be used for gene mapping cloning