Zheng Xiaojian, Gong Yongchang, Kumar Dhiraj, Chen Fei, Kuan Sulan, Liang Zi, Hu Xiaolong, Cao Guangli, Xue Renyu, Gong Chengliang
School of Biology and Basic Medical Science, Soochow University, Suzhou, 215123, China.
National Engineering Laboratory for Modern Silk, Soochow University, Suzhou, 215123, China.
Transgenic Res. 2016 Aug;25(4):507-16. doi: 10.1007/s11248-016-9947-3. Epub 2016 Mar 7.
Bombyxin (BBX) is an insulin-like peptide exists in the silkworm Bombyx mori. Our previous studies on the effects of inhibiting BBX-B8 expression found that BBX-B8 is important for the development of organ, reproduction and trehalose metabolism in the silkworms. In this paper, we investigated the expression profile of the BBX-B8 gene and effect of BBX-B8 overexpression on the development, body weight, silk protein synthesis and egg diapause of B. mori to further understand BBX-B8 functions. BBX-B8 gene expression could be detected in the brains, midguts, anterior silkglands, ovaries, testes, fat bodies, hemolymph, malpighian tubules and embryos by RT-PCR, however it was mainly expressed in the brain. Western blots showed that the change in BBX-B8 expression was not obvious in the brain of 1- to 4-day-old larvae of fifth instar silkworms, but expression increased substantially at 5- to 6-day-old larvae of fifth instar silkworms. Transgenic silkworms overexpressing BBX-B8 were obtained by introducing non-transposon transgenic vector pIZT-B8 containing a BBX-B8 gene driven by Orgyia pseudotsugata nucleopolyhedrovirus IE2 promoter into the genome. Development duration of the transgenic silkworms was delayed by 2.5-3.5 days. Cocoon shell weight of transgenic silkworms was reduced by 4.79 % in females and 7.44 % in males, pupal weight of transgenic silkworms was reduced 6.75 % in females and 13.83 % in males compared to non-transgenic silkworms, and 5.56-14.29 % of transgenic moths laid nondiapausing eggs. All results indicated that BBX-B8 plays an important role in the development, silk protein synthesis and egg diapause of silkworm.
家蚕素(BBX)是一种存在于家蚕(Bombyx mori)体内的类胰岛素肽。我们之前关于抑制BBX - B8表达影响的研究发现,BBX - B8对家蚕的器官发育、生殖和海藻糖代谢很重要。在本文中,我们研究了BBX - B8基因的表达谱以及BBX - B8过表达对家蚕发育、体重、丝蛋白合成和卵滞育的影响,以进一步了解BBX - B8的功能。通过RT - PCR可在家蚕的脑、中肠、前部丝腺、卵巢、睾丸、脂肪体、血淋巴、马氏管和胚胎中检测到BBX - B8基因表达,但其主要在脑中表达。蛋白质免疫印迹显示,五龄1至4日龄幼虫脑中BBX - B8表达变化不明显,但在五龄5至6日龄幼虫中表达大幅增加。通过将含有由云杉卷叶蛾核多角体病毒IE2启动子驱动的BBX - B8基因的非转座子转基因载体pIZT - B8导入基因组,获得了过表达BBX - B8的转基因家蚕。转基因家蚕的发育历期延迟了2.5 - 3.5天。与非转基因家蚕相比,转基因家蚕的雌茧层重降低了4.79%,雄茧层重降低了7.44%;转基因家蚕的雌蛹重降低了6.75%,雄蛹重降低了13.83%,并且5.56 - 14.29%的转基因蛾产下非滞育卵。所有结果表明,BBX - B8在家蚕的发育、丝蛋白合成和卵滞育中起重要作用。