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BAC selection of the large white butterfly, Pieris brassicae (Lepidoptera: Pieridae) containing orthologs of the silkworm, Bombyx mori (Lepidoptera: Bombycidae)

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Abstract

We constructed a bacterial artificial chromosome (BAC) library from the large white butterfly, Pieris brassicae (L.) (Lepidoptera: Pieridae), which composed of 15,360 clones with an average insert size of 103 kb. We designed 39 degenerate oligonucleotide primer pairs and cloned the PCR products for sequencing. From these plus pierisin data, we designed single tagged sequence primers and selected 40 BAC clones using a 3-step PCR protocol. Additional 40 BACs were sequenced to improve identification of P. brassicae BACs carrying orthologs of the silkworm, Bombyx mori (L.) (Lepidoptera: Bombycidae). Among 34 BAC clones containing multiple B. mori orthologs, sequences in 27 BACs correspond to a block (less than 1 Mb) of every B. mori chromosome. While orthologs in 5 BACs showed collinearity to each B. mori chromosome with a distance (1.4–6.8 Mb), orthologs in two BACs corresponded to genes in more than two different B. mori chromosomes.

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Acknowledgements

This study was supported by JSPS grant numbers, JP16H05050, JP19H02972 and, JP19K06067.

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Authors and Affiliations

  1. Division of Biocontrol and Bioenvironmental Sciences, United Graduate School of Agricultural Sciences, Iwate University, Ueda 3-18-8, Morioka, 020-8550, Japan

    Mizuki Ohno, Toshiaki Fujimoto & Ken Sahara

  2. Faculty of Agriculture, Iwate University, Ueda 3-18-8, Morioka, 020-8550, Japan

    Toshiaki Fujimoto & Ken Sahara

  3. Faculty of Agriculture, Hokkaido University, N9, W9, Kita-ku, Sapporo, 060-8550, Japan

    Yota Naito

  4. Agricultural Research Institute, HOKUREN Federation of Agricultural Cooperatives, Naganuma, 069-1317, Japan

    Yota Naito

  5. Institute of Agrobiological Sciences, National Agricultural and Food Research Organization, Owashi 1-2, Tsukuba, 305-8634, Japan

    Akiya Jouraku & Yuji Yasukochi

Authors
  1. Mizuki Ohno
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  2. Toshiaki Fujimoto
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  3. Yota Naito
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  4. Akiya Jouraku
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  5. Yuji Yasukochi
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  6. Ken Sahara
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Correspondence to Ken Sahara.

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Ohno, M., Fujimoto, T., Naito, Y. et al. BAC selection of the large white butterfly, Pieris brassicae (Lepidoptera: Pieridae) containing orthologs of the silkworm, Bombyx mori (Lepidoptera: Bombycidae). Appl Entomol Zool 55, 159–174 (2020). https://doi.org/10.1007/s13355-019-00665-7

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  • DOI: https://doi.org/10.1007/s13355-019-00665-7

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