State Key Laboratory of Silkworm Genome Biology, Key Laboratory for Sericulture Biology and Genetic Breeding, Ministry of Agriculture and Rural Affairs, Southwest University, Chongqing 400716, China; Chongqing Engineering and Technology Research Center for Silk Biomaterials and Regenerative medicine, Southwest University, Chongqing 400716, China.
State Key Laboratory of Silkworm Genome Biology, Key Laboratory for Sericulture Biology and Genetic Breeding, Ministry of Agriculture and Rural Affairs, Southwest University, Chongqing 400716, China; Chongqing Engineering and Technology Research Center for Silk Biomaterials and Regenerative medicine, Southwest University, Chongqing 400716, China.
Int J Biol Macromol. 2020 Jun 1;152:525-534. doi: 10.1016/j.ijbiomac.2020.02.296. Epub 2020 Feb 26.
NPC1 is a large glycoprotein with 13 transmembrane-spanning domains, which plays a crucial biological role in cholesterol transport and metamorphosis of animals. However, the physiological functions of this gene have rarely been elucidated in insects. Here, we isolated the NPC1 gene from Bombyx mori (BmNPC1), sequenced and evaluated its physiological functions. BmNPC1 comprised of 3702 bp open reading frame, encoding a protein of 1233 amino acid residues. The recombinant protein was expressed, and anti-BmNPC1 antibodies were synthesized. Immunofluorescence assay revealed that BmNPC1 protein localized in the cytoplasm of the cells. The qRT-PCR analysis showed that BmNPC1 expression was most significant in the testis, followed by the malpighian tubules, hemocytes, and ovary. The knockdown of BmNPC1 by double-stranded RNA caused the accumulation of cholesterol in the cells. Furthermore, suppression of this gene influenced the expression of ecdysone-responsive genes and also prevented the molting in B. mori (Dazao) larvae. Overall, BmNPC1 may have different biological roles in the physiology of silkworm, B. mori (Dazao), since it regulates the cholesterol transport and molting process.
NPC1 是一种具有 13 个跨膜结构域的大型糖蛋白，在胆固醇运输和动物变态中起着至关重要的生物学作用。然而，该基因在昆虫中的生理功能很少被阐明。在这里，我们从家蚕（BmNPC1）中分离出 NPC1 基因，对其进行测序并评估其生理功能。BmNPC1 由 3702bp 的开放阅读框组成，编码 1233 个氨基酸残基的蛋白质。表达了重组蛋白，并合成了抗 BmNPC1 抗体。免疫荧光分析显示 BmNPC1 蛋白定位于细胞质中。qRT-PCR 分析表明，BmNPC1 在睾丸中的表达最显著，其次是马氏管、血细胞和卵巢。双链 RNA 敲低 BmNPC1 导致细胞内胆固醇积累。此外，该基因的抑制还影响蜕皮激素反应基因的表达，并阻止家蚕（大造）幼虫蜕皮。总体而言，BmNPC1 在家蚕（大造）的生理过程中可能具有不同的生物学作用，因为它调节胆固醇运输和蜕皮过程。
