TY - JOUR T1 - Tuatara (Sphenodon) genomics: BAC library construction, sequence survey, and application to the DMRT gene family JF - J Hered Y1 - 2006 A1 - Wang, Z. A1 - Miyake, T. A1 - Edwards, S. V. A1 - Amemiya, C. T. KW - *Chromosomes, Artificial, Bacterial KW - *Multigene Family KW - Animals KW - Bayes Theorem KW - Gene Library KW - genome KW - Genomics/methods KW - Male KW - Phylogeny KW - Reptiles/*genetics KW - Transcription Factors/classification/*genetics AB -

The tuatara (Sphenodon punctatus) is of "extraordinary biological interest" as the most distinctive surviving reptilian lineage (Rhyncocephalia) in the world. To provide a genomic resource for an understanding of genome evolution in reptiles, and as part of a larger project to produce genomic resources for various reptiles (evogen.jgi.doe.gov/second_levels/BACs/our_libraries.html), a large-insert bacterial artificial chromosome (BAC) library from a male tuatara was constructed. The library consists of 215 424 individual clones whose average insert size was empirically determined to be 145 kb, yielding a genomic coverage of approximately 6.3x. A BAC-end sequencing analysis of 121 420 bp of sequence revealed a genomic GC content of 46.8%, among the highest observed thus far for vertebrates, and identified several short interspersed repetitive elements (mammalian interspersed repeat-type repeats) and long interspersed repetitive elements, including chicken repeat 1 element. Finally, as a quality control measure the arrayed library was screened with probes corresponding to 2 conserved noncoding regions of the candidate sex-determining gene DMRT1 and the DM domain of the related DMRT2 gene. A deep coverage contig spanning nearly 300 kb was generated, supporting the deep coverage and utility of the library for exploring tuatara genomics.

VL - 97 N1 -

0022-1503 (Print)Journal ArticleResearch Support, N.I.H., ExtramuralResearch Support, U.S. Gov't, Non-P.H.S.

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