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Cloning, Expression, and Cell Localization of a Novel Small Heat Shock Protein Gene: BmHSP25.4

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Abstract

Using molecular approaches, a new member of the Bombyx mori small heat shock protein family was cloned and characterized. The isolated gene contains an open reading frame of 672 bp, encodes a polypeptide of 223 amino acid residues with a predicted molecular mass of 25.4 kDa, and is therefore named BmHSP25.4. The gene codes for a protein homologous to the previously characterized HSP20.4 and HSP19.9. Western blotting analysis revealed that BmHSP25.4 existed in the fifth-instar larva’s fatty body and blood tissues. Immunohistochemistry assay also showed that BmHSP25.4 was located in the fifth-instar larva’s fatty body. The results of above studies have indicated constitutive expression of BmHSP25.4 in fatty body, blood tissues, and Bm5 cells. Finally, we examined the effect of heat stress on localization of BmHSP25.4 using anti-BmHSP25.4 polyclonal antibody by immunofluorescence. Under normal conditions, BmHSP25.4 was mostly found in the cytoplasm. However, after heat treatment, most of BmHSP25.4 distributed in the cell membrane. After 3 h of recovery following the heat shock treatment, the localization of BmHSP25.4 was the same as that under normal conditions.

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Acknowledgements

This work was supported by financial grants from the National High Technology Research and Development Program (no. 2007AA021703, no. 2007AA100504) and the National Basic Research Program of China (no. 2005CB121006).

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  1. Yaozhou Zhang

    Present address: Institute of Biochemistry, Zhejiang Sci-Tech University, Xiasha High-Tech Zone No.2 Road, Hangzhou, 310018, China

Authors and Affiliations

  1. Institute of Biochemistry, Zhejiang Sci-Tech University, Hangzhou, 310018, China

    Qing Sheng, Jiayin Xia & Zuoming Nie

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  1. Qing Sheng
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  2. Jiayin Xia
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  4. Yaozhou Zhang
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Correspondence to Yaozhou Zhang.

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Sheng, Q., Xia, J., Nie, Z. et al. Cloning, Expression, and Cell Localization of a Novel Small Heat Shock Protein Gene: BmHSP25.4 . Appl Biochem Biotechnol 162, 1297–1305 (2010). https://doi.org/10.1007/s12010-009-8890-7

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  • DOI: https://doi.org/10.1007/s12010-009-8890-7

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