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3758939
Genetic Engineering
Descripción
Basics of Genetic Engineering
Sin etiquetas
biology
engineering
molecular
molecular bioengineering
masters
Mapa Mental por
Alex Schiffer7426
, actualizado hace más de 1 año
Más
Menos
Creado por
Alex Schiffer7426
hace más de 8 años
40
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Resumen del Recurso
Genetic Engineering
Genome Modification
CRISPR
CAS9 cuts dsDNA
Uses guide RNA
Least specific/ Easiest to program
TALEN
34 AAs, 12&13 recognize Nucleotides
Recruits FOK1 to cut dsDNA
Zinc-finger
Most Specific / Hardest to Program
Each Module Recognizes 3 bp
Recruits FOK1 (Cuts dsDNA)
NHEJ/HEJ - Lambda Red Phage
FLP Recombinase Recognizes FRT
Flank Insert Gene With FRT Sites
FLP Expression Plasmid Eliminates Antibiotic Resistance Gene
Exo, Gamma, Beta
MAGE
Alters Many Loci at the Same Time
Add Pool of Short Oligos that Create Mismatch/Insertion/Deletion
Various Mutants Created
CAGE
Prokaryotic Gene Regulation
RNA Polymerase
Holoenzyme = Core + Sigma
Core - Transcribes
Sigma - Initiates
1st Level of Transcriptional Regulation
Regulate Large Numbers of Genes
Transcription Factors
2nd Level of Transcriptional Regulation
Regulates a Few Genes
Lac Operon
Lactose Metabolism
Lactose - Disaccharide Sugar
Lactose Permease
Membrane Protein
Transport Lactose Into Cell
β-Galactosidase
Cleaves Lactose
Diauxic Shift
Small Plateau After Glucose Consumed to Switch to Lactose Usage
Carbon Catabolite Repression
Presence of Preferred Carbon Source Prevents Use of Secondary Carbon Source
CCR in E.coli
CAP Protein
Lactose Activates Transcription of β-Galactosidase
LacZ
β-Galactosidase Mutation
LacY
Permease Mutation
LacA
Transacetylase Mutation
LacI
Upstream Mutation
Creates a Repressor
Eukaryotic Gene Regulation
Six Types of Regulation
Transcriptional Control
Chromatin Mediated
Mediators
Epigenetic Control
Math
Gibbs Free Energy
When a Reaction will Occur
Laws of Thermodynamics
1st Law
2nd Law
Boltzmann Distribution
Molecular Cloning
Traditional Method
Digest, Ligate, Transform
Uses Restriction Enzymes
Gibson Method
T5 Exonuclease Chews Back Homologous 5' Ends
DNA Fragments Anneal to Each Other
Phusion Polymerase Extends 3' Ends
Taq Ligase Seals Nicks
Doesn't Have to Be Palindromic (Avoids Poisoning)
Phage λ Recombination
Gateway System
In-Fusion Method
Golden Gate One Pot Digestion Ligation
Modular Cloning
Sequencing
Sanger
Next Generation
Nucleic Acids
DNA
Bases
C-G (3 H-bonds)
A-T (2 H-bonds)
in RNA, T is replaced with U
Pyrimidines
C, U, T (smaller)
Only Anti
Purines
A & G (bigger)
Anti or Syn
Replication
Semi-Conservative
Enzymes
RNA Primase
DNA Polymerase
DNA Helices
DNA Ligase
Topoisomerase
Repair
Recognize Errors
Exonuclease
MutS & MutL
Single Strand Repair
BER
DNA Glycosylase
Endonuclease
Phosphodiesterase
DNA Polymerase
DNA Ligase
NER
Excision Nuclease
DNA Helicase
DNA Polymerase
DNA Ligase
Double Strand Repair
NHEJ
Section of DNA removed
Causes Frame Shift
HEJ
Removes Damaged DNA
Uses Sister Chromatid to Repair
Types
B-DNA
B'-DNA
A-DNA
Z-DNA
RNA
mRNA
tRNA
rRNA
Glycosidic Bonds
Phosphodiester Bonds
Sugars
C2'-Endo (S)
C3'-Endo (N)
Proteins
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