BIOL 200 Study Guide - Final Guide: T7 Rna Polymerase, Rna Polymerase Iii, Dna-Binding Domain
10 Feb 2016
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Hybridize oligo-t primers to mrna, use reverse transcriptase to transcribe cdna. Attach poly dg adapter to 3 prime end, use poly dc primer, dna pol synthesizes complimentary strand. How does lac operon work for increasing transcription. Iptg activates lac promoter, activating gene for t7 rna pol, making t7 rna polymerase. T7 rna pol binds to t7 promoter located in front of target gene, transcribing sequence of interest like crazy! Crystallography and x ray diffraction helps show what a protein really looks like in biological context. You can get the crystal, blast it with an x ray beam, you get diffraction pattern. This gives you an idea of the electron density of the crystals. You can then fill structure in with known amino acids. Allows you to put together an amino acid chain, and try to work on building together a final structure. Small kd = proteins with a strong affinity for each other.
