Separation Of The Silkworm, Bombyx Mori Natural Membrane Protein Bmb019518 Of Purification

Membrane proteins, especially integral membrane proteins, play a very significant role in material and energy exchanges between the cell and the outside circumstance. Various membrane proteins exist as channel proteins or carrier proteins and are an important channel in keeping the inter circumstance steadily. Membrane protein is also important in regulating cell's metabolism, even in the electron transmit chain. Membrane proteins possess 1/3 of cell total proteins, and most of them are the target of new medicine. It is helpful and beneficial to study on the membrane proteins for the medicine develop and human health.Integral membrane proteins exist in a lipid environment of biological membranes, which results in a difficult work to purify them with traditional method. In this article, we use the material of silkworm pupa infected by BmNPV, and peel the host cell membrane via the Baculoviral budding. Using homogenate, differential centrifuge, density gradient centrifuge, high pressure crush and high speed centrifuge. The high quality membrane was obtained, and then was extracted with CHAPS to dissolve these membrane proteins. The solved membrane proteins were further isolated with ion-exchange chromatograph and size exclusion chromatograph using the AKTA explorer10 protein purified system. We analysed these proteins with 2D electrophoresis. After digging the sepecial protein dots and digested them with trypsin, we obtained the protein's PMF (Peptide Mass Fingerprinting) through the MALDI-TOF-MS. The PMF datas were analysed with the software of PLGS Browser. It was finally confirmed that a transmembrane protein was obtained, which had one transmembrane domain. The ID of this membrane protein in the Southwest University Protein Data Bank is Bmb019518. That the Baculoviral Budded Viruses were used to obtain high quality membrane protein offered a novel method for isolating membrane proteins, and lay a good foundation for the further studies on the structure and function of the membrane proteins.