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

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• 作者：Jianqing Chen ; Jian Chen ; Qijing Gai ; Zhengbing Lv ; Dan Wang ; Zuoming Nie ; Jiang Wang ; Xuedong Wang ; Xiangfu Wu and Yaozhou Zhang
• 关键词：Troponin C ; Development ; Expression pattern ; Immunohistochemistry ; Bombyx mori (Insecta)
• 刊名：Cell and Tissue Research
• 出版年：2008
• 出版时间：March, 2008
• 年：2008
• 卷：331
• 期：3
• 页码：725-738
• 全文大小：821.7 KB
• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Biomedicine
  Human Genetics
  Proteomics
  Molecular Medicine
  
• 出版者：Springer Berlin / Heidelberg
• ISSN：1432-0878

文摘

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.