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Construction and characterization of a normalized cDNA library from the river snail Bellamya aeruginosa after exposure to copper

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Abstract

The construction of a normalized cDNA library is a popular tool for identifying novel biomarkers for monitoring environmental pollution. In the present study, a normalized cDNA library was constructed from the river snail Bellamya aeruginosa after exposure to Cu2+ by using the SMART technique. The titer of the cDNA library was 1.78 × 106 pfu/ml, with a recombinant efficiency of 95.8%. In addition, from 6,000 randomly selected and sequenced clones, 5,473 high-quality ESTs were identified. After processing the sequences, 3,961 unigenes representing 897 contigs and 3,064 singlets were obtained with 27.6% redundancy. Analysis of expressed sequenced tags using COG and GO annotation and KEGG pathway data showed that a large group of genes related to growth and development, signal transduction, and defense mechanisms were present in the cDNA library. Based on our findings, this normalized cDNA library will provide a valuable resource for further research on functional genes and ecotoxicology in B. aeruginosa.

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Abbreviations

ESTs:

Expressed sequenced tags

COG:

Clusters of orthologous groups of proteins

GO:

Gene ontology

DSN:

Duplex-specific nuclease

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Acknowledgments

Financial supports by National Basic Research Program of China Grant (2007CB407306), Water Pollution Control and Management (2009ZX07528-003) and China’s new environmental problems (2009467109) are gratefully acknowledged.

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Authors and Affiliations

  1. State Key Laboratory of Environmental Criteria and Risk Assessment, Chinese Research Academy for Environment Sciences, Beijing, 100012, China

    Zi-Cheng Li, Li-Hui An, Ying Liu, Lei Zhang, Hao Chen, Xing-Ru Zhao, Bing-Hui Zheng & Lin-Bo Zhang

  2. State Environmental Protection Key Laboratory of Estuarine and Coastal Research, Chinese Research Academy for Environment Sciences, Beijing, 100012, China

    Zi-Cheng Li, Li-Hui An, Qing Fu, Ying Liu, Lei Zhang, Hao Chen, Xing-Ru Zhao, Li-Jing Wang & Bing-Hui Zheng

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  1. Zi-Cheng Li
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  2. Li-Hui An
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  3. Qing Fu
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  4. Ying Liu
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  5. Lei Zhang
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  6. Hao Chen
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  10. Lin-Bo Zhang
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Correspondence to Li-Hui An.

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Li, ZC., An, LH., Fu, Q. et al. Construction and characterization of a normalized cDNA library from the river snail Bellamya aeruginosa after exposure to copper. Ecotoxicology 21, 260–267 (2012). https://doi.org/10.1007/s10646-011-0786-y

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