分子实验报告2010030038生05陶欣

The summer experiment——the Molecular PartBio05 陶欣 2025030038Group 8-2 Class BExperiment partner: 刘子源 金雨2025/8/24-2025/8/28I.ObjectivesThe overall objective of this summer all-around experiment is to improve our experimental operation skills and problem-designing skills, to build up our good habits in working and study, to learn how to cooperate with our partners, and to obtain a certain level of standard writing ability through the process of the expression, purification and identification of the HSP16.3 protein.II.Background and Principles1. Small heat shock protein (sHSP)In response to environmental stresses, cells increase the expression of one class of important proteins called small heat shock proteins (sHSP), which endow cells with thermo-tolerance and are believed to be involved in the inhibition of apoptosis, organization of the cytoskeleton, establishing the refractive properties of the eys lens, and stabilization of RNA.Proteins of sHSP family have various biological functions, but all of them share the ATP independent chaperone activity, preventing protein aggregation in stress. Structure studies indicate that they usually function by assembling into certain oligo-structure.2. HSP16.3HSP16.3 is an important sHSP from Mycobacterium Tuberculosi, it is markedly expressed during the stationary phase and becomes a main protein in the cell. HSP16.3 is an immunodominant antigen and the major membrane protein of M.tuberculosis. Previous studies showed that HSP16.3 can make the cell structure stable and prevent stationary M.tuberculosis from autolysing. Gene knockout studies indicate that HSP16.3 is required for M.tuberculosis to grow in macrophages.HSP16.3 consists of 144 amino acids and its molecular weight is 162...