Memorias do Instituto Oswaldo Cruz (Oct 2006)

A bacterial artificial chromosome library for Biomphalaria glabrata, intermediate snail host of Schistosoma mansoni

  • Coen M Adema,
  • Mei-Zhong Luo,
  • Ben Hanelt,
  • Lynn A Hertel,
  • Jennifer J Marshall,
  • Si-Ming Zhang,
  • Randall J DeJong,
  • Hye-Ran Kim,
  • David Kudrna,
  • Rod A Wing,
  • Cari Soderlund,
  • Matty Knight,
  • Fred A Lewis,
  • Roberta Lima Caldeira,
  • Liana K Jannotti-Passos,
  • Omar dos Santos Carvalho,
  • Eric S Loker

DOI
https://doi.org/10.1590/S0074-02762006000900027
Journal volume & issue
Vol. 101
pp. 167 – 177

Abstract

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To provide a novel resource for analysis of the genome of Biomphalaria glabrata, members of the international Biomphalaria glabrata Genome Initiative (biology.unm.edu/biomphalaria-genome.html), working with the Arizona Genomics Institute (AGI) and supported by the National Human Genome Research Institute (NHGRI), produced a high quality bacterial artificial chromosome (BAC) library. The BB02 strain B. glabrata, a field isolate (Belo Horizonte, Minas Gerais, Brasil) that is susceptible to several strains of Schistosoma mansoni, was selfed for two generations to reduce haplotype diversity in the offspring. High molecular weight DNA was isolated from ovotestes of 40 snails, partially digested with HindIII, and ligated into pAGIBAC1 vector. The resulting B. glabrata BAC library (BG_BBa) consists of 61824 clones (136.3 kb average insert size) and provides 9.05 × coverage of the 931 Mb genome. Probing with single/low copy number genes from B. glabrata and fingerprinting of selected BAC clones indicated that the BAC library sufficiently represents the gene complement. BAC end sequence data (514 reads, 299860 nt) indicated that the genome of B. glabrata contains ~ 63% AT, and disclosed several novel genes, transposable elements, and groups of high frequency sequence elements. This BG_BBa BAC library, available from AGI at cost to the research community, gains in relevance because BB02 strain B. glabrata is targeted whole genome sequencing by NHGRI.

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