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ARS Home » Southeast Area » Stoneville, Mississippi » Warmwater Aquaculture Research Unit » Research » Publications at this Location » Publication #142808

Title: Construction and characterization of a BAC library from a gynogenetic channel catfish, Ictalurus punctatus

item Quiniou, Sylvie
item MILLER, N
item WILSON, M
item Wolters, William
item Waldbieser, Geoffrey - Geoff

Submitted to: Genetics Selection Evolution
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 4/9/2003
Publication Date: 11/1/2003
Citation: Quiniou, S., Katagiri, T., Miller, N.W., Wilson, M., Wolters, W.R., Waldbieser, G.C. 2003. Construction and characterization of a BAC library from a gynogenetic channel catfish, Ictalurus punctatus. Genetics Selection Evolution. 35(6):673-683.

Interpretive Summary: Marker-assisted selection can be used to increase the efficiency of catfish broodstock selection, but genetic maps are currently limited. In order to facilitate gene identification and development of catfish genetic maps, channel catfish genomic DNA was divided into DNA fragments maintained as bacterial artificial chromosome (BAC) clones. The average size of a genomic DNA fragment in the BAC library was 165,000 bases, and the library contained enough clones for a 7-fold coverage of the catfish genome. These clones will be useful for development of linkage and physical genetic maps for channel catfish, and for the identification of genes controlling important production traits.

Technical Abstract: Two bacterial artificial chromosome (BAC) libraries, designated CCBL1 and CCBL2, were constructed by cloning Hind III-digested high molecular weight DNA from a gynogenetic channel catfish, Ictalurus punctatus, into the vector pBeloBAC11. Approximately 53,500 clones from CCBL1 were arrayed in 384-well plates and stored at -80 degrees Celsius, while CCBL2 was amplified and stored at -80 degrees Celsius without arraying. Pulsed-field gel electrohoresis of 100 clones after Not I 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 CCBL1 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.