CHEM 451 Study Guide - Final Guide: Dna Shuffling, Oligomer, Crystallin
Document Summary
Dna shuffling = randomly recombining dna orthologs from diff species for the same protein (homologous but not identical) to insert into plasmid for expression. > fragment with nucleus, melta dna, allow to reanneal, extend sections. > primerless pcr (assume enough homology between diff regions so they"ll reanneal) > sequence final library to check it differs from parental sequence. Random mutagenesis + dna shuffling optimises proteins we want shsp. Form oligomers, ring structures (storage form, not active form) Heavily conserved alpha crystallin fold within monomer subunits for ring structure assembly. > n & c terminal seq of fold gives variability/ specificity, n terminal seq evolves indep. > folded n terminus important for intra ring interactions/ form interface with ring below it. When cellular stress -> oligomer disassembles to functional subunits (dimers, tetramers) Regulated by modifying n terminal region for oligomer -> functional dimeric subunit. Hsp16. 9 = 12 monomer subunits, 2x6 rings stacked on top of each other.
