Nie Zuoming, Xu Jiangtao, Chen Jian, Lv Zhengbing, Wang Dan, Sheng Qing, Wu Yi, Wang Xuedong, Wu Xiangfu, Zhang Yaozhou
Institute of Biochemistry, College of Life Sciences, Zhejiang Sci-Tech University, Hangzhou, China.
Appl Biochem Biotechnol. 2009 Jul;158(1):59-71. doi: 10.1007/s12010-008-8302-4. Epub 2008 Jul 17.
A recombinant Bombyx mori profilin protein (rBmPFN) was overexpressed in Escherichia coli BL21. Purified rBmPFN was used to generate anti-BmPFN polyclonal antibody, which were used to determine the subcellular localization of BmPFN. Immunostaining indicated that profilin can be found in both the nucleus and cytoplasm but is primarily located in the cytoplasm. Real-time RT-PCR and Western blot analyses indicated that, during the larvae stage, profilin expression levels are highest in the silk gland, followed by the gonad, and are lowest in the fatty body. Additionally, BmPFN expression begins during the egg stage, increases during the larvae stage, reaches a peak during the pupa stage, and decreases significantly in the moth. Therefore, we propose that BmPFN may play an important role during larva stage development, especially in the silk gland.
一种重组家蚕肌动蛋白结合蛋白(rBmPFN)在大肠杆菌BL21中过表达。纯化后的rBmPFN用于制备抗BmPFN多克隆抗体,该抗体用于确定BmPFN的亚细胞定位。免疫染色表明,肌动蛋白结合蛋白在细胞核和细胞质中均有发现,但主要位于细胞质中。实时RT-PCR和蛋白质印迹分析表明,在幼虫阶段,肌动蛋白结合蛋白在丝腺中的表达水平最高,其次是性腺,在脂肪体中的表达水平最低。此外,BmPFN的表达始于卵期,在幼虫期增加,在蛹期达到峰值,而在蛾期显著下降。因此,我们认为BmPFN可能在幼虫阶段发育过程中发挥重要作用,尤其是在丝腺中。