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Construction of cDNA subtractive library from pearl oyster (Pinctada fucata Gould) with red color shell by SSH

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Abstract

The molecular basis of color polymorphism in the shells of the pearl oyster Pinctada fucata is largely unknown. We developed a red-shelled family line and used suppression subtractive hybridization (SSH) to screen for differentially expressed genes in red- and non-red-shelled pearl oysters. We constructed forward and reverse cDNA subtractive libraries consisting of 2 506 and 797 clones, respectively. Among 343 randomly selected clones in the forward library, 304 sequences were identified in GenBank using BLASTx and BLASTn. Of the 304 sequences, 13 showed no similarity to known sequences and 291 were matched with known genes of the pearl oyster, including shematrin-1, shematrin-2, shematrin-6, shematrin-7, nacrein, nacrein-like protein, aspein for shell matrix protein, glycine-rich protein, mantle gene 5, 28S, EST00031, EST00036, 16S, and COI. In the reverse library, 7 clones were sequenced and analyzed by BLAST. Two sequences shared similarity with EST00036 from the P. fucata subtraction cDNA library, four with the P. fucata mitochondrial gene for 16S rRNA and 1 with P. fucata shematrin-2. We evaluated the expression of 12 genes from the forward library using RT PCR. Two sequences matched with 16S and COI so were considered to be false positives. The remaining 10 sequences were differentially expression in the red-shelled pearl oysters. Our results suggest that differential expression of these genes may be related to color variation in the red-shelled family line of the pearl oyster.

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Authors and Affiliations

  1. Key Laboratory of Marine Bio-resources Sustainable Utilization, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou, 510301, China

    Yunyan Guan  (管云雁), Liangmin Huang  (黄良民) & Maoxian He  (何毛贤)

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  1. Yunyan Guan  (管云雁)
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  2. Liangmin Huang  (黄良民)
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  3. Maoxian He  (何毛贤)
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Correspondence to Maoxian He  (何毛贤).

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Supported by National High Technology Research and Development Program of China (863 Program) (No. 2006AA10A409), the Knowledge Innovation Program of the South China Sea Institute of Oceanology, Chinese Academy of Sciences (No. SQ200906), the Science and Technology Program of Guangdong Province (No. 2008A020100004), the National Key Technology Research and Development Program (No. 2007BAD29B01-8), and the Key Plan for Marine Development of Guangdong Province (No. A200708C01)

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Guan, Y., Huang, L. & He, M. Construction of cDNA subtractive library from pearl oyster (Pinctada fucata Gould) with red color shell by SSH. Chin. J. Ocean. Limnol. 29, 616–622 (2011). https://doi.org/10.1007/s00343-011-0200-x

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  • DOI: https://doi.org/10.1007/s00343-011-0200-x

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