<p> Agro/Ansc/Bio/Gene/Hort 305 Fall, 2017 Review Sheet for Exam #3</p><p>TRANSLATION (Chapter 13): </p><p>Overview, the genetic code, the Wobble hypothesis; Basic structure of amino acids and the nature of their side chains (R group); peptides, peptide bonds, peptidyl transferase, primary, secondary, tertiary and quaternary structures of proteins; function of proteins. tRNA: structure: anticodon and amino acid acceptor arm; aminoacyl tRNA synthetase, charging tRNA, coding recognition, wobble in anticodon. Ribosomes: small subunit, large subunit, rRNA, ribosomal proteins, the E,P, and A sites on the small subunit. Decoding of mRNA: polyribosomes, (polysomes), initiation codon, initiation of translation, Shine-Dalgarno sequence, initiation factors, charged tRNAs; peptidyl transferase. Translation termination, release factors, stop codons. Translation in eukaryotes: The role of Cap, scanning of 5’UTR, differences in translation initiation between eukaryotes and prokaryotes. Focus on the following concepts: Differences and similarities in translation between eukaryotes and prokaryotes Coupled transcription and translation in prokaryotes The role of aminoacyl tRNA synthetase and peptidyl transferase</p><p>REGULATION OF GENE EXPRESSION IN PROKARYOTES (CHAPTER 14): </p><p>Overview of the common points of regulation. Transcriptional Regulation: Operons – cis-acting elements, promoters, activator binding sites, operators, terminators, structural genes; regulatory gene, repressor protein.</p><p>Polycistronic RNA. </p><p>Effector molecules: inducer, co-repressor, inhibitors, allosteric shifts Positive regulation, negative regulation. Induction, repression, activation. Constitutive genes, inducible and repressible operons.</p><p>The lactose (lac) operon: Regulation in the presence of lactose and glucose; role cAMP; the effect of mutations on the different elements of the operon and in the regulatory gene. The tryptophan (trp) operon: regulation by repression and Attenuation; the unique features of the leader sequence (Trpl)</p><p>Translational control: RNA binding repressor, antisense RNA.</p><p>Post-translational regulation: feed-back inhibition/end product inhibition, allosteric enzymes, covalent modification.</p><p>Effect of mutations in the different elements in the lac and Trp operons </p><p>REGULATION OF GENE EXPRESSION IN EUKARYOTES (Chapters 15, 16):</p><p>Overview of the levels of regulation.</p><p>Transcriptional regulation: </p><p>Core promoter and Regulatory elements - enhancers, silencers. Properties of enhancer and silencer elements.</p><p>Transcription factors (TF): General/ basal transcription factors, specialized transcription factors - activators and repressor; TFIID, Mediator Mechanism by which the regulatory transcription factors regulate transcription.</p><p>Chromatin structure and gene expression: ATP-dependent chromatin remodeling complex. Histone acetyl transferase and histone deacetylase – their role in chromatin compaction</p><p>DNA methylation: its role and mechanism by which it works in gene silencing</p><p>Alternate splicing: outcome, sliceosomes, splice sites, splicing factors miRNA mRNA stability: length of poly (A)-tail, destabilizing sequences in the 3’UTR</p><p>Translational control: Role of initiation factors, RNA binding proteins, regulation at the level of translation and RNA stability of iron regulated genes (iron response elements, iron response elements – ferritin and transferin receptor.</p><p>RECOMBINANT DNA TECHNOLOGY (Chpt 20) Gene Cloning: Cloning vectors : selectable markers, origin of replication, cloning site, LacZ : their function with regards to cloning.</p><p>Restriction endonuclease: Restriction sites, cohesive or sticky ends, blunt ends. Ligations, transformation cDNA synthesis: reverse transcriptase.</p><p>Polymerase chain reaction (PCR) : Design of primers, the three steps involved in each cycle, Taq polymerase </p>