BmOsi 9a基因在家蚕胚胎细胞周期中的功能研究

The Functional Study of the BmOsi 9a Gene in the Cell Cycle of Silkworm Embryonic Cells

Osiris基因家族是一类昆虫特有且进化保守的基因家族。已知Osi 9亚家族基因在鳞翅目昆虫中发生特异扩增,产生高度同源的6个拷贝,且表达模式也具有同步性,但该基因家族扩增机制和生物学功能未知。为了揭示BmOsi 9a基因的生物学功能,本研究通过荧光定量分析和Western blot实验检测到BmOsi 9a在家蚕胚胎细胞(BmE)中有表达,免疫荧光分析发现BmOsi 9a主要存在于细胞质中,构建过表达及dsRNA干涉载体,结果显示过表达对细胞的表型和增殖没有显著影响,而转染干涉载体下调BmOsi 9a表达则使细胞的增殖受到显著影响。利用流式细胞技术分析发现G0/G1期细胞显著增加,S期细胞则显著减少,表明BmOsi 9a与BmE细胞的增殖相关。研究结果表明BmOsi 9a在细胞周期过程发挥作用,为进一步阐明BmOsi 9a生物学功能奠定了基础。

The Osiris gene family is an evolutionarily conserved and insect-specific gene family.The Osi9 genes undergoes specific duplication in Lepidoptera insects,producing six highly homologous members with synchronized expression.However,the duplication mechanism and biological function of this gene family are currently unknown.To elucidate the biological function of BmOsi 9a gene,BmOsi 9a was detected to be expressed in silkworm embryonic cells(BmE)through fluorescence quantitative detection and Western blot experiments.Immunofluorescence analysis revealed that BmOsi 9a is primarily located in the cytoplasm.Overexpression and dsRNA interference vectors for BmOsi 9a were constructed.Overexpression of BmOsi 9a did not show a significant effect on cell phenotype and proliferation,while transfection with the interference vector targeting BmOsi 9a significantly affected cell proliferation.Flow cytometry analysis showed a significant increase in G0/G1 phase cells and a significant decrease in S phase cells,indicating BmOsi 9a should be involved in cell proliferation.These results indicated that BmOsi 9a plays a role in the cell cycle,laying a foundation for further elucidating the biological functions of BmOsi 9a.