感染BmNPV家蚕幼虫马氏管蛋白质组的初探.pdfVIP

感染BmNPV 家蚕幼虫马氏管蛋白质组的初探 1 1，2* 1，2 1 1，2 1，2 王娜 唐顺明 裘智勇 丁丽平 赵巧玲 沈兴家 1 2 （江苏科技大学，江苏镇江212018 ； 中国农业科学院蚕业研究所，农业部蚕桑遗传改良重点开放实验室，江苏 镇江212018 ） 摘 要 本文旨在探讨家蚕幼虫马氏管的蛋白质组及其免疫功能。我们构建了重组BmNPV-EGFP，注射 感染家蚕，调查病毒在各组织感染时相（以呈现EGFP为指标）；以呈现EGFP的马氏管为材料，采用双向电 泳（2-DE）技术对其蛋白组进行了分离与差异分析。结果表明，病毒在各组织感染时相先后为血细胞（72 hpi），后部丝腺（72 hpi），气管（84 hpi），脂肪体组织（84 hpi）,马氏管（96 hpi），中肠（96 hpi）； 试验组100 μg蛋白经双向电泳及银染后可以检测出551个蛋白点，而对照组样品检测出472个蛋白点，存 在明显差异，说明病毒感染后马氏管蛋白组有所变化；利用ImageMaster 2D Platinum软件找到了4个表 达明显上调的蛋白点，供进一步用基质辅助激光解析电离飞行时间质谱法（MALDI-TOFMS）鉴定。 关键词 家蚕；马氏管；BmNPV；双向电泳；蛋白质组 Preliminary research on proteomics of Malpighian tuble from the silkworm Bombyx mori infected with BmNPV WANG Na1 TANG Shun-Ming1,2* QIU Zhi-Yong1,2 DING Li-Ping1 ZHAO QIAO-Ling1,2 SHEN Xing-Jia1,2 1 2 Jiangsu University of Science and Technology, Zhenjiang Jiangsu 212018，China; The Key Laboratory of Silkworm and Mulberry Genetic Improvement ，Ministry of Agriculture ，Sericultural Research Institute ，Chinese Academy of Agricultural Sciences ，Zhenjiang Jiangsu 212018 ，China Abstract ：In this paper, we preliminary investigate the proteomics and immune function of malpighian tubule of the silkworm. At first, the recombinant BmNPV-EGFP was constructed, and its infection process in the larvae tissue was determined. The EGFP is firstly appeared in hemolymph and posterior silk gland at 72 hpi , then in trachea and fat body at 84 hpi , and lastly in malpighian tubule and midgut 96 hpi . Then, we apply the method of two-dimensional polyacrylamide gel electrophoresis 2-DE to investigate the comparative proteomics of malpighian tubule which infected with BmNPV-EGFP or not. The results showed that there are 551 spots in the BmNPV group, while only 472 spots in the control group, suggesting that the infection of BmNPV resulted in the change of proteomics of malpighian tubule. Lastly, 4 spots that increased dramatically in the infected one was excised and supplied to identify furtherly by means of matrix assisted la