MCB 2050 Study Guide - Final Guide: Dna Ligase, Expression Vector, Lac Operon
9 Dec 2018
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Chapter 14 summary and mcb 2050 review 1-4. Spread transformants on agar with x-gal: blue colonies (lacz intact no insert), white colonies (lacz gene disrupted insert present). For isolation and amplification of dna sequences. Contains (i) origin of replication, (ii) selectable marker, (iii) mcs. Example: pbluescript (also has blue/white selection for plasmids without/with inserts) For the expression of proteins in bacteria, yeast, plant or animal cells. Contains promoter appropriate for target cell located upstream of mcs. Also contains elements found in cloning vector (for cloning/amplification in e. coli) Eukaryotic expression plasmids must contain a promoter upstream of the mcs and a polya signal downstream of the mcs. Example: pcmv, a mammalian cell expression vector. For the measurement of gene regulatory sequences. Contains a reporter gene that when expressed can be quantitatively measured. Mcs is located upstream of the reporter gene. Also contains elements found in a cloning vector. Libraries can be screened by complementation or hybridization with specific probes.
