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首页> 外文期刊>Cell and Tissue Research >Molecular characterization and immunohistochemical localization of a novel troponin C during silkworm development.
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Molecular characterization and immunohistochemical localization of a novel troponin C during silkworm development.

机译：家蚕发育过程中新型肌钙蛋白C的分子表征和免疫组化定位。

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摘要

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]X(6)E, a highly conserved putative Ca(2+) -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.
机译：我们已经克隆并测序了一个新的家蚕基因，该基因编码与其他已知的肌钙蛋白C（TnC）蛋白质具有高度同源性的蛋白质。 BmTnC中也存在氨基酸序列DX [DN] X [DSG] X（6）E，这是一种高度保守的推定Ca（2+）结合基序，该基序存在于先前鉴定的TnC蛋白球状结构域的环中。我们已经表达并纯化了具有大约21.6 kDa分子量的带有His标记的BmTnC融合蛋白。我们已经使用这种纯化的融合蛋白来生产抗BmTnC的多克隆抗体，用于蛋白质印迹分析。这些分析表明，BmTnC在幼虫头部，马氏小管，表皮，睾丸和肠道中表达，这已通过免疫组织化学证实。此外，实时逆转录/聚合酶链反应显示，这些组织之间的BmTnC mRNA水平存在显着差异。我们的发现表明，BmTnC在家蚕的肌肉组织中选择性表达，包括头部，马氏小管，体壁和肠道的一部分。

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