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| Prohibitin (PHB) is an evolutionarily conserved ubiquitously expressed multifunctional protein. However, its mechanism of action is unknown as yet. To better characterize the prohibitin protein from silkworm pupae (BmPHB), a gene encoding a prohibitin protein was identified from a cDNA library of silkworm pupae, which has an ORF of 825 bp, encoding a predicted 274 amino acids. A His-tagged BmPHB fusion protein was expressed in Escherichia coli Rosetta (DE3) and purified with affinity and reversed-phase chromatographies. Purified rBmPHB was used to generate anti-BmPHB polyclonal antibody, which was used to determine the subcellular localization of BmPHB. Immunostaining indicated that prohibitin can be found in both nucleus and cytoplasm but is located primarily in cytoplasm. Western blot analyses indicated that, in the fifth instar larva, BmPHB was expressed descendingly in gonad, malpighian tubule, trachea, fatty body, intestine and head. However, none was detected in larva’s silk gland and epidermis. Additionally, BmPHB was expressed in the nascent egg, larva and pupa, but none was detected in the moth. Therefore, we hypothesized that BmPHB may play an important role on silkworm development. This is a first description of the prohibitin protein from silkworm pupae, B. mori. |
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| Keywords:Bombyx mori;Prohibitin;Expression analysis;Subcellular localization |
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