Proteomic Analysis Of The Silkworm Fat Body During Metamorphosis And The Interaction Between Hemolin And Yippee Protein

Silkworm(Bombyx mori) is an important model insect of Lepidoptera, an ideal material for the study of metamorphosis development. Fat body is a kind of important tissue organ in the body, it is not only the center of metabolism and hormone, but also play an extremely important role in the growth of silkworm development. In order to find the protein which is closely related to the development of metamorphosis, we carried out the study on the proteomics of silkworm fat body during metamorphosis. In addition, according to the development of insect wing is a typical process of metamorphosis, we studied the interaction of Hemolin protein and Yippee protein expressed in the wing disc, which provides the basis for the study of the development process in the future. The results of this study are as follows:1. We isolated and identified 27 differentially expressed protein spots by 2-DE technique from the total protein of fifth instar larvae and pupa in different developmental stages; Under the analysis of mass spectrometric and database search, we matched the candidate protein. These proteins are closely related to the material and energy metabolism, cytoskeleton, metabolism and protein synthesis, growth development, signal transduction and physiological activities, they also play important function and role in silkworm growth and metamorphosis development.2. 9 protein spots were selected from 27 differential protein spots, including, 70 K heat shock protein, prolyl hydroxylase, elongation factor, glutathione solution transferase, succinyl coenzyme A, pancreatic chymotrypsin inhibitor, cytoplasmic formyl tetrahydrofolate dehydrogenase and beta N-acetyl Glucosaminidase, and the corresponding genes were detected by fluorescence quantitative analysis. The possible roles of them in abnormal development were also analyzed.3. Polyclonal antiserum was prepared to study the interaction between Hemolin protein and Yippee protein. The polyclonal antiserum against Yippee protein was prepared by using New Zealand white rabbits. The results of Blotting Western showed that the polyclonal antiserum of the silkworm Yippee protein has good specificity which can meet the demand of our experiment.4. We carried out Co-IP experiment of Hemolin and Yippee protein using the two polyclonal antiserum we prepared. The results showed that the Hemolin and Yippee proteins interact with each other in the wing disc.Through the above research, we found some important proteins which are related to the metamorphosis, and we believe that the study has some reference value to the research of metamorphosis. The study of the interaction between Hemolin and Yippee provides some clues for the development of the wing disc.