State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing, China.
College of Biotechnology, Southwest University, Chongqing, China.
Biochem Biophys Res Commun. 2020 Feb 5;522(2):492-498. doi: 10.1016/j.bbrc.2019.11.094. Epub 2019 Nov 25.
Sericin, produced in the middle silk gland (MSG) of silkworms, is a group of glue proteins that coat and cement silk fibers. Several genes are known to encode sericin, but their spatiotemporal regulation has yet to be fully elucidated. Here, we report in detail the expression profiles of the promoters of two major sericin-coding genes, Sericin 1 (Ser1)and Sericin 3 (Ser3), by analyzing Gal4/UAS transgenic silkworms. We found that UAS-linked EGFP fluorescence in transgenic silkworms driven by Ser1-Gal4was detected in only the R3, R4 and R5 regions of MSG starting inday-3 fifth-instar larvae and was continuously expressed until silk gland degradation. In transgenic silkworms driven by Ser3-Gal4, EGFP fluorescence was detected at a low level in the R2 region of MSG since the last day of fifth-instar larvae, and the expression increased during the wandering stages and was continuously detected until silk gland degradation. The molecular detection of EGFP expression in each of the Gal4/UAS transgenic silkworms was consistent with fluorescence observations. These findings reveal clear differences in the regulatory characteristics of the promoters of Ser1and Ser3 and provide new insights into the regulatory mechanism of the expression of sericin-coding genes.
丝胶是由家蚕中部丝腺(MSG)产生的一组胶状蛋白,它包裹并黏合丝纤维。已知有几个基因编码丝胶,但它们的时空调控尚未完全阐明。在这里,我们通过分析 Gal4/UAS 转基因家蚕,详细报告了两个主要丝胶编码基因 Sericin 1(Ser1)和 Sericin 3(Ser3)启动子的表达谱。我们发现,由 Ser1-Gal4 驱动的转基因家蚕中 UAS 连接的 EGFP 荧光仅在 3 龄第 5 天幼虫的 MSG 的 R3、R4 和 R5 区域检测到,并持续表达直到丝腺降解。由 Ser3-Gal4 驱动的转基因家蚕中,EGFP 荧光在 MSG 的 R2 区域从 5 龄幼虫的最后一天开始以低水平检测到,在游走阶段表达增加,并持续检测到丝腺降解。Gal4/UAS 转基因家蚕中 EGFP 表达的分子检测与荧光观察结果一致。这些发现揭示了 Ser1 和 Ser3 启动子的调控特征的明显差异,并为丝胶编码基因表达的调控机制提供了新的见解。