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Novel Isoforms of N16 and N19 Families Implicated for the Nacreous Layer Formation in the Pearl Oyster Pinctada fucata

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Abstract

Although a wide variety of proteins and genes possibly related to the shell formation in bivalve have been identified, their functions have been only partially approved. We have recently performed deep sequencing of expressed sequence tags (ESTs) from the pearl oyster Pinctada fucata using a next-generation sequencer, identifying a dozen of novel gene candidates which are possibly associated with the nacreous layer formation. Among the ESTs, we focused on three novel isoforms (N16-6, N16-7, and N19-2) of N16 and N19 families with reference to five known genes in the families and determined the full-length cDNA sequences of these isoforms. Reverse transcription-polymerase chain reaction revealed that N16-6 was expressed in gill, gonad, adductor muscle, and mantle, whereas N16-7 exclusively in mantle. N19-2 was expressed in all tissues examined. In situ hybridization demonstrated their regional expression in mantle and pearl sac, which well corresponded to those shown by EST analysis previously reported. Shells in the pearl oyster injected with dsRNAs of N16-7 and N19-2 showed abnormal surface appearance in the nacreous layer. Taken together, novel isoforms in N16 and N19 families shown in this study are essential to form the nacreous layer.

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Authors and Affiliations

  1. Graduate School of Agricultural and Life Sciences, The University of Tokyo, Bunkyo-ku, Tokyo, 113-8657, Japan

    Fumito Ohmori, Shigeharu Kinoshita, Shuichi Asakawa & Shugo Watabe

  2. Graduate School of Bioresources, Mie University, Kurimamachiya 1577, Tsu, Mie, 514-8507, Japan

    Daisuke Funabara

  3. Graduate School of Biosphere Science, Hiroshima University, Hiroshima, 739-8528, Japan

    Hiroki Koyama

  4. Pearl Research Laboratory, K. Mikimoto & Co., Ltd., Osaki Hazako 923, Hamajima, Shima, Mie, 517-0403, Japan

    Kiyohito Nagai

  5. Mikimoto Pharmaceutical Co., Ltd., Kurose 1425, Ise, Mie, 516-8581, Japan

    Kaoru Maeyama & Kikuhiko Okamoto

  6. School of Marine Biosciences, Kitasato University, Minami-ku, Sagamihara, Kanagawa, 252-0313, Japan

    Shugo Watabe

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  1. Fumito Ohmori
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  2. Shigeharu Kinoshita
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  3. Daisuke Funabara
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  6. Kaoru Maeyama
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  7. Kikuhiko Okamoto
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  8. Shuichi Asakawa
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  9. Shugo Watabe
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Correspondence to Shugo Watabe.

Electronic supplementary material

Supplementary Table 1

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Supplementary Table 2

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Supplementary Table 3

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Supplementary Fig. 1

The DNA nucleotide sequences of N16 isoforms, N16–1, N16–2, N16–3, N16–5, N16–6 and N16–7, from the pearl oyster Pinctada fucata. (PPTX 60.4 kb)

Supplementary Fig. 2

The DNA nucleotide sequences of N19 isoforms, N19 and N19–2, from the pearl oyster Pinctada fucata. (PPTX 61.2 kb)

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Ohmori, F., Kinoshita, S., Funabara, D. et al. Novel Isoforms of N16 and N19 Families Implicated for the Nacreous Layer Formation in the Pearl Oyster Pinctada fucata. Mar Biotechnol 20, 155–167 (2018). https://doi.org/10.1007/s10126-017-9793-1

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  • DOI: https://doi.org/10.1007/s10126-017-9793-1

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