Characterization of a gene encoding prohibitin in silkworm, Bombyx mori

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• 作者：Zhengbing Lv^a ; ^b ; Xufang Zhang^a ; ^b ; Lili Liu^a ; Jian Chen^a ; ^b ; Zuoming Nie^a ; ^b ; Qing Sheng^a ; ^b ; Wenping Zhang^a ; ^b ; Caiying Jiang^a ; ^b ; Wei Yu^a ; ^b ; Dan Wang^a ; ^b ; Xiangfu Wu^a ; ^b ; Shijun Zhang^c ; ^d ; Jun Li^d ; Yaozhou Zhang^a ; ^b ; ^{yaozhou@zstu.edu.cn}
• 关键词：PHB1 ; Prohibitin 1 ; PHB domain ; Prohibitin-like domain ; DAPI ; 4&rsquo ; -6-Diamidino-2-phenylindole ; PVDF ; polyvinylidene fluoride
• 刊名：Gene
• 出版年：2012
• 出版时间：10 July, 2012
• 年：2012
• 卷：502
• 期：2
• 页码：118-124
• 全文大小：871 K

文摘

Background

Prohibitin (PHB) is an evolutionarily conserved multifunctional protein with ubiquitous expression. However, its molecular roles are largely unknown.

Methods

To better understand the function of prohibitin protein in silkworm (BmPHB), its coding sequence was isolated from a cDNA library of silkworm pupae. An His-tagged BmPHB fusion protein was expressed in Escherichia coli Rosetta (DE3) and purified with affinity and reversed-phase chromatography. Purified rBmPHB was used to generate anti-BmPHB polyclonal antibody. The subcellular localization of BmPHB was analysed by immunohistochemistry.

Results

BmPHB gene has an ORF of 825 bp, encoding a predicted peptide with 274 amino acid residues. Immunostaining indicate that prohibitin is expressed in nucleus and predominately 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, no expression was detected in larva's silk gland and epidermis. In addition, BmPHB was expressed in the nascent egg, larva and pupa, but not in the moth.

Conclusions

The expression of BmPHB gene presents differential characteristic in different stage and tissues. It may play important roles in the development of silkworm.

General significance

Studies on prohibitin have been still restricted to a few specific insects and insect cell lines such as Drosophila, Acyrthosiphon pisum and mosquito cell lines, not yet in silkworm. This is a first characterization of prohibitin in silkworm, B. mori.