Genet. Sel. Evol.
Volume 35, Number 6, November-December 2003
|Page(s)||673 - 683|
Construction and characterization of a BAC library from a gynogenetic channel catfish Ictalurus punctatusSylvie M.A. Quinioua, b, Takayuki Katagiria, Norman W. Millera, Melanie Wilsona, William R. Woltersb and 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)
A bacterial artificial chromosome (BAC) library was constructed by cloning HindIII-digested high molecular weight DNA from a gynogenetic channel catfish, Ictalurus punctatus, into the vector pBeloBAC11. 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 NotI 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.
Key words: bacterial artificial chromosome / catfish / genome / Ictalurus punctatus
Correspondence and reprints: Geoffrey C. Waldbieser firstname.lastname@example.org
© INRA, EDP Sciences 2003