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Molecular characterization and immunohistochemical localization of a novel troponin C during silkworm development

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Abstract

We have cloned and sequenced a novel Bombyx mori gene that encodes a protein having a high degree of homology with other known troponin C (TnC) proteins. The amino acid sequence, DX[DN]X[DSG]X6E, a highly conserved putative Ca2+-binding motif found in loops within the globular domains of previously identified TnC proteins, is also present in BmTnC. We have expressed and purified to homogeneity a His-tagged BmTnC fusion protein having a molecular weight of approximately 21.6 kDa. We have used this purified fusion protein to produce polyclonal antibodies against BmTnC for Western blot analyses. These analyses have revealed that BmTnC is expressed in the larval head, the Malpighian tubule, the epidermis, the testis, and the gut, as has been confirmed by immunohistochemistry. In addition, real-time reverse transcription/polymerase chain reaction has shown that BmTnC mRNA levels differ substantially among these tissues. Our findings indicate that BmTnC is selectively expressed in the muscular tissues of the silkworm, including portions of the head, the Malpighian tubule, the body wall, and the gut.

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Authors and Affiliations

  1. Institute of Biochemistry, Zhejiang Sci-Tech University, 2nd Street, Xiasha Higher Education Zone, Hangzhou, 310018, China

    Jianqing Chen, Jian Chen, Qijing Gai, Zhengbing Lv, Dan Wang, Zuoming Nie, Jiang Wang, Xuedong Wang, Xiangfu Wu & Yaozhou Zhang

  2. Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, Shanghai, 200031, China

    Xiangfu Wu

Authors
  1. Jianqing Chen
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  2. Jian Chen
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  3. Qijing Gai
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  4. Zhengbing Lv
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  5. Dan Wang
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  6. Zuoming Nie
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  7. Jiang Wang
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  8. Xuedong Wang
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  9. Xiangfu Wu
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  10. Yaozhou Zhang
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Correspondence to Yaozhou Zhang.

Additional information

This work was supported by grants from the National High Technology Research and Development Program of China (no. 2005AA206120) and the National Basic Research Program of China (no. 2005CB121006).

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Chen, J., Chen, J., Gai, Q. et al. Molecular characterization and immunohistochemical localization of a novel troponin C during silkworm development. Cell Tissue Res 331, 725–738 (2008). https://doi.org/10.1007/s00441-007-0516-1

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  • DOI: https://doi.org/10.1007/s00441-007-0516-1

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