Genetics - DNA
Terms
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- One copy of a gene
- Allele
- A segment of DNA made of distinct nucleotide pairs on a certain location on a chromosome that codes for a specific protein
- Gene
- Purines/Pyrimidines
- -A and G (2ring) -C and T (1ring)
- DNA Melting Point Equation
- T = (G+C)4 + (A+T)2
- GCN4
- like Polymerase but can read sequence without unwinding
- Substitution Inversion Translocation
- - another nucleotide - flipping around of sequence - part of one chromosome moves to another
- Protein Structure
- -determined by sequence of amino acids -primary -secondary - a and B -tertiary - myoglobin
- Replace a base
- Base Analogs
- Amino Acid Changes - explain
- Samesense-codon substitution Missense-change sequence Nonsense-stop codons stop protein synthesis Frameshift-insert or delete nucleotides
- Collection of mutations that do not compliment each other (on separate genes)
- Complementation Group
- Virus that infects bacteria cells
- Bacteriophage (T4 - affects E.Coli)
- Gene Conversion
- correction of heteroduplex DNA to other sequence
- Polypeptide
- molecular chain of amino acids
- DNA Mutation
- change in nucleotide sequence at a particular position
- Recombination in a gene
- causes wild-type
- Deletion Mapping
- determines location of mutations
- Multimer with identical subunits
- Homodimer
- Protein
- substance from amino acids joined by peptide bonds
- Components of a Gene
- -Regulatory Region - promoter -Coding Region - forms products -Termination Region - tells polymerase to stop transcription
- Location of mutations can be determined by:
- Deletion mapping and Fine structure mapping
- Mapping studies showed that genes are located in:
- Linear stretches of DNA
- Amino acid sequence:
- Directs the folding of a polypeptide to secondary and tertiary structures
- Exon and Intron
- Exon-coding sequence Intron-removed after transcription
- Sequence of nucleotides that specify the sequence of amino acids in a polypeptide
- Genetic Code
- Genetic Code features
- -code is degenerate -consists of codons -reading frame is initiated by start codon -Code reads 5'-3'
- Where Transcription and Translation take place
- Transcription-nucleus Translation-cytoplasm
- Copying DNA to single stranded RNA
- Transcription
- What is needed for transcription?
- Promoter, RNA polymerase
- Replication Fork
- moves up double helix
- 2 Things Needed for Transcription
- Promoter and RNA Polymerase
- Splicesome
- protein complex that performs splicing -5 snRNA's (small nuclear RNA) and Proteins
- Lariat
- 2'-5' phosphodiester bond formed during splicing in middle of RNA
- Alternative Splicing
- produces 2 different mRNA's from same gene
- Translation Elongation
- site to site transfer and amino acid bond formation
- Glycosylation
- sugars are added to protein
- Intragenic Suppression
- when 1BP is deleted, 1BP is inserted
- Cloning
- using a complex mixture of restriction fragments in cells to make many exact replicas one fragment at a time
- 4^n Rule
- gives approximate size of restriction fragments
- Vectors
- agents that carry recombinant DNA to foreign genes or DNA
- Plasmids
- hold restriction sites for cloning -Drug resistance marker - selects bacteria with plasmid -Origin of Replication
- LacZ
- bacterial gene that encodes for an enzyme beta-galactosidase
- Screening Bacteria
- -Isolate bacteria with plasmid DNA -Alkaline Lysis-mix DNA with Ethidium Bromide and Cesium Chloride -Centrifuge to separate plasmid from Genomic DNA
- Library
- bacteria that contain vectors with different inserts of entire genome
- Genomic Library
- contains parts of genes
- cDNA
- contains coding information only
- Expression Vectors
- used to express protein in bacteria
- Colony Hybridization
- determines which bacterial colony contains the cloned DNA
- Southern Blotting
- Transferring DNA to nitrocellulose filter through capillary action
- PCR
- Polymerase Chain Reaction -amplification of target DNA sequence -Needs - TAQ polymerase, 5'-'3 primers, Buffer, dNTP's