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Characterization of AFLAV, a Tf1/Sushi retrotransposon from Aspergillus flavus

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Abstract

The plasmid, pAF28, a genomic clone from Aspergillus flavus NRRL 6541, has been used as a hybridization probe to fingerprint A. flavus strains isolated in corn and peanut fields. The insert of pAF28 contains a 4.5 kb region which encodes a truncated retrotransposon (AfRTL-1). In search for a full-length and intact copy of retrotransposon, we exploited a novel PCR cloning strategy by amplifying a 3.4 kb region from the genomic DNA of A. flavus NRRL 6541. The fragment was cloned into pCR®4-TOPO®. Sequence analysis confirmed that this region encoded putative domains of partial reverse transcriptase, RNase H, and integrase of the predicted retrotransposon. The two flanking long terminal repeats (LTRs) and the sequence between them comprise a putative full-length LTR retrotransposon of 7799 bp in length. This intact retrotransposon sequence is named AFLAV (A. flavus Retrotransposon). The order of the predicted catalytic domains in the polyprotein (Pol) placed AFLAV in the Tf1/sushi subgroup of the Ty3/gypsy retrotransposon family. Primers derived from AFLAV sequence were used to screen this retrotransposon in other strains of A. flavus. More than fifty strains of A. flavus isolated from different geological origins were surveyed and the results show that many strains have extensive deletions in the regions encoding the capsid (Gag) and Pol.

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Author notes

  1. Alice S. Tarun

    Present address: Seattle Biomedical Research Institute, 307 Westlake Ave N Suite 500, Seattle, WA, 98109, USA

  2. Sonal N. Pandey

    Present address: Applied Biosystems, 850 Lincoln Centre Drive, Foster City, CA, 94404, USA

Authors and Affiliations

  1. U.S. Department of Agriculture, Agricultural Research Service, Western Regional Research Center, 800 Buchanan Street, Albany, CA, 94710, USA

    Sui-Sheng T. Hua, Alice S. Tarun & Leo Chang

  2. MJ Bioworks, Inc., South San Francisco, CA, USA

    Sonal N. Pandey

  3. U.S. Department of Agriculture, Agricultural Research Service, Southern Regional Research Center, New Orleans, LA, USA

    Perng-Kuang Chang

Authors
  1. Sui-Sheng T. Hua
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  2. Alice S. Tarun
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  3. Sonal N. Pandey
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  4. Leo Chang
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  5. Perng-Kuang Chang
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Correspondence to Sui-Sheng T. Hua.

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Hua, SS.T., Tarun, A.S., Pandey, S.N. et al. Characterization of AFLAV, a Tf1/Sushi retrotransposon from Aspergillus flavus . Mycopathologia 163, 97–104 (2007). https://doi.org/10.1007/s11046-006-0088-8

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  • DOI: https://doi.org/10.1007/s11046-006-0088-8

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