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Sequence analysis and expression of a cDNA clone encoding tropomysin in Sinonovacula constricta

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Abstract

Shellfish can cause severe anaphylactic reactions. Tropomyosin has been assumed partly responsible for the cross-reactivity among shellfish and other invertebrates. In this study, cDNA of Sinonovacula constricta was amplified by RT-PCR and 3′-RACE from total RNA. The obtained tropomyosin cDNA included an open reading frame coding for 284 amino acids. The deduced amino acid sequence of the corresponding protein shared high identity with other allergenic tropomyosins. Expression of the recombinant tropomyosin was carried out in Escherichia coli BL21(DE3) using vector PET28a and the purification of the recombinant protein was performed via affinity chromatography. IgE reactivity of recombinant tropomyosin was investigated by immunoblot and the sensized precentage was 36% which indicated that tropomyosin was the minor allergens in S. constricta. Moreover, the character of the purified protein was analyzed by MALDI-TOF-MS.

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Acknowledgments

Authors thank Mr. Yong Wang for the MALDI-TOF-MS analysis and the Shenzhen people’s hospital for kindly sera supporting. This study was supported by National 863 high technology research (No. 2006AA100308).

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Authors and Affiliations

  1. Institute of Allergy and Immunology, Shenzhen University, Shenzhen, 518060, China

    Juanjuan Song, Li Li, Zhigang Liu & Qiyuan Li

  2. Guangzhou Medical College, Guangzhou, 510182, China

    Juanjuan Song & Pixin Ran

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  1. Juanjuan Song
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  2. Li Li
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  3. Zhigang Liu
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  4. Qiyuan Li
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  5. Pixin Ran
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Correspondence to Zhigang Liu.

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Juanjuan Song and Li Li contributed equally to this work.

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Song, J., Li, L., Liu, Z. et al. Sequence analysis and expression of a cDNA clone encoding tropomysin in Sinonovacula constricta . Mol Biol Rep 36, 315–321 (2009). https://doi.org/10.1007/s11033-007-9181-4

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  • DOI: https://doi.org/10.1007/s11033-007-9181-4

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