关键词: 家蚕, 生存素, 组织表达； H3Ser10ph, 有丝分裂, BmN4细胞

Abstract: 【Aim】 To clarify the function of survivin gene in the mitosis of BmN4 cells of Bombyx mori. 【Methods】 The expression levels of BmSurvivin in different tissues (silk gland, midgut, Malpighian tubules, testis, ovary, fat body, epidermis and cuticle) of the day-3 5th instar larvae of B. mori were detected by qRT-PCR. The fusion vector pIZT/V5-His-BmSurvivin-GFP (BG) was constructed and transfected into BmN4 cells. The localization of BmSurvivin and Ser10 phosphorylated histone H3 (H3Ser10ph) in different mitotic stages of BmN4 cells was detected by immunofluorescence. 【Results】 The expression level of BmSurvivin was the highest in the Malpighian tubules of the day-3 5th instar larvae of B. mori, followed by that in the silk gland and midgut. pIZT/V5-His-BmSurvivin-GFP vector was successfully constructed. The immunofluorescence results showed that obvious GFP signal could be seen in the interphase nucleus of BmN4 cells, covering the nuclear and cytoplasmic regions indicating the location of BmSurvivin. At the mitotic phase, BmSurvivin was co-localized with chromatin indicated by GFP signal. When the BmN4 cells formed obvious double orientation, GFP signal was also visible in the spindle region. At the anaphase stage, with the separation of sister chromatids, BmSurvivin was located in the chromatin and cytokinesis region indicated by GFP signal. At the telophase stage, with the formation of two new daughter cells, BmSurvivin was located only in the cytokinesis region indicated by GFP signal. H3Ser10ph was located at the site of chromatin condensation at the prometaphase stage of BmN4 cells, and the signal intensity was the strongest at the metaphase, coinciding with the whole chromosome. The signal disappeared at the anaphase stage. 【Conclusion】 BmSurvivin is associated with the dynamic changes of chromatin and spindle during mitotic cycle.

Key words: Bombyx mori, survivin, tissue expression, H3Ser10ph, mitosis, BmN4 cell