Ligation of multiple DNA fragments through uracil-DNA glycosylase ...
DNA Repair. Transcription Differences Between RNA & DNA 1.) RNA has 2’ OH 2.) RNA has uracil...
-
Upload
harold-oliver -
Category
Documents
-
view
232 -
download
3
Transcript of DNA Repair. Transcription Differences Between RNA & DNA 1.) RNA has 2’ OH 2.) RNA has uracil...
DNA Repair
Transcription
Differences Between RNA & DNA
1.) RNA has 2’ OH2.) RNA has uracil instead of thymine3.) RNA is single stranded (but can fold to self base-pair)
RNA Polymerases--- catalyze the polymerization of RNA using a
DNA template--- in prokaryotes RNA polymerases require a sigma
factor to bind DNA and initiate transcription
Sigma Factors--- a protein that binds a specific nucleotide sequence,
called a promoter--- a bacterium will generally have multiple sigma factors
(allows for better gene regulation)--- one sigma factor usually controls most of the central
metabolic and biosynthesis genes ( 70 in E. coli)
Transcription
Regulation of Transcription
1.) Sigma factors
2.) Other DNA binding proteins
A.) Negative, block RNA polymerase’s access to DNA--- lac repressor
B.) Positive, enhance recruitment of RNA polymerase
--- catabolite activation
--- sometimes both negative and positive systems compete to operate on the same set of genes
--- genes in bacteria are often grouped into regulatory elementscalled operons (groups of genes controlled by the same operator),this saves time and energy for the cell
Lactose Operon in E. coli
Tryptophan Operon
Positive Regulation: The Maltose Operon
Catabolite Activation, dual regulation: (positive & negative) in the lac operon
Two Component Systems
Translation
Requires:1.) Ribosomes2.) charged tRNAs3.) mRNA4.) accessory factors (initiation factor,
elongation factor, etc.)5.) ATP & GTP
Based on Genetic Code
--- need at least 20 “words” so must have 3 nucleotide codons--- code is degenerate, multiple codons specify the same
amino acid, effects:A.) reduces likelyhood of mutational damageB.) codon useage can vary between different
organisms
Translation
Other Tidbits:
RNA interference:
--- Use non-coding (ncRNA) to block translation or trigger destruction of mRNA
Quorum Sensing:
--- Small rapidly diffusing molecules are used to measurecell density
--- Often used by pathogens to time toxin release formaximum effect
Mutation
--- a change in the genetic information (DNA sequence) of an organism
--- cells spend considerable energy fixing mutations--- preventing all mutation would actually be a bad idea, WHY?
Types of Mutation:
A.) Base Substitution: swap one base for another
1.) Transition: change from one purine or pyrimidine base to another (AG or TC)
2.) Transversion: change from a purine to a pyrimidine or a pyrimidine to a purine
--- only possibility of change to AA sequence of protein, WHY?
Types of Mutations (Continued):
B.) Insertion/ Deletion: add or remove one or more base pairs
--- these types of mutations can cause a frameshift, changing the reading frame of the rest of the gene (often results in a prematurestop codon).
--- What is one case where there would be almost no effectof a frameshift mutation?
Mutagens and Mutagenesis
Mutagen: something that causes DNA damage
1.) Chemicals, chemical modification of DNA bases caneasily result in mutation
2.) DNA intercalators, aromatic compounds that canform stacking interactions with bases, areoften “read” as additional bases, creatinginsertion mutations
3.) UV- induced thymine dimers
4.) Ionizing radiation, creates double stranded DNAbreaks, can be difficult to repair and may stillresult in an insertion or deletion
--- mutagenesis is the process of purposely mutating an organism,“wreck and check” strategy of protein study
The Ames Test:
--- measuring how mutagenic a substance is