Micro 10 Transcription and Translation
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Chantelsham on October 10, 2009
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104 terms
Terms | Definitions |
|---|---|
 Transcription DNA -> RNA the 1st step to making | protein. |
| The single strand of RNA that results from reading the DNA is called | messenger RNA (mRNA) |
| 1st step RNA polymerase binds to a promoter | is just a codon that says "start here" |
| T T C G A G C A | DNA |
| A A G C U C G U | RNA |
| Transcription RNA polymerase binds to the | promoter |
| RNA Transcription, Moves down the strand of | mRNA reading each base and building RNA's complimentary base. |
| RNA Transcription Arrives at a STOP signal and | detaches. |
| RNA Transcription | Result is a single strand of mRNA |
| Translation mRNA -> | protein |
| Translation, Ribosomal RNA (rRNA) binds to the | mRNA |
| Ribosomal RNA | rRNA |
| Translation, The rRNA "translates" the mRNA into an | amino acid sequence |
| The entire process of transcription and translation is are referred to as | gene expression. |
| Gene expression, Every gene ultimately results in the production of | protein. |
| Phenotype | The physical appearance or functional expression of a trait. (blonde hair, green eyes, estrogen receptor) |
| Genotype | The genetic code underlying a single trait or set of traits. (specific DNA sequence for blonde hair, green eyes, estrogen receptor) |
| Regulating Gene Expression, How are transcription and translation controlled | 60-80% of genes are constitutive |
| 60-80% of genes are constitutive and | Produced at a fixed rate |
| How are transcription and translation controlled | Two mechanisms control gene expression Repression & Induction |
| Two mechanisms control gene expression | Repression & Induction |
| Repression Inhibits | Transcription |
| Repression is Usually due to a | build up of end-product |
| Repression , As the end-product builds up, repressors bind to the | promoter region of DNA and prevent RNA polymerase from binding |
| Transcription is turned on by an | inducer. |
| Enzymes that require an inducer to be produced are called | inducible enzymes |
| Lactose acts as an inducer for | b-galactosidease (ONPG) |
| b-galactosidease breaks lactose into | glucose and galactose. |
| Mutations are changes in the | hereditary message of an organism. |
| Two categories of mutations | Sequence changes & Changes in Gene position |
| Sequence changes | Base substitution, Chemical modification, DNA breaks, Slipped mispairing , Triplet expansion |
| Changes in Gene position | Chromosomal rearrangement & Insertional inactivation |
| Sequence Changes | Base substitution, Normal C G T C A, Base sub C G G C A, Occurs during replication, Also called a point mutation |
| Sequence Changes | Chemical modification, A base becomes chemically modified by a mutagenic chemical, Another class of point mutation |
| Sequence changes DNA | breaks |
| Sequence changes Caused by | Ionizing radiation |
| Sequence changes Causes | double strand breaks |
| Sequence changes Results in the | loss (deletion) of short segments |
| Sequence changes Slipped | Mispairing |
| Sequence changes Results in | a deletion |
| Sequence changes Also called a | frameshift mutation |
| Sequence changes, A sequence is present in | more than one place on a strand and pairs out of order |
| Sequence changes Like a shirt buttoned on the | wrong button |
| Triplet Expansion is When a | 3-base sequence is repeated several times in a gene. |
| Triplet Expansion Results in | mutant proteins |
| More than 15 human diseases are attributed to | triplet expansion |
| Mutations - Changes in Gene Position | Chromosomal Rearrangement |
| Translocation | Part of one chromosome becomes a part of another chromosome |
| Inversion | Orientation of a portion of a chromosome is reversed |
| Duplication (gene abnormalities) | Entire regions of chromosome become duplicated |
| Deletion(gene abnormalities) | Similar to the deletion on a nucleotide level but much larger amounts of DNA are deleted |
| Aneuploidy (gene abnormalities) | Whole chromosomes are lost or gained |
| Polyploidy(gene abnormalities) | An entire SET of chromosomes are added. |
| Genetic Recombination Technology, Modern scientists can insert or delete gene segments to | form new combinations of genes |
| Genetic Recombination Technology | Genes are usually inserted |
| Genetic Recombination Technology, Vaccine development is trying to delete | pathogenic genes. |
| What do the letters PCR stand for | Polymerase, Chain, Reaction |
| PCR is the amplification of a | DNA sequence |
| PCR is the name of the assay that scientists use to look at what genes are being expressed in a | cell, tissue, or organism. |
| Why amplification? We have not yet developed the technology to | detect 1 copy of DNA although it is fast approaching. |
| Orientation to DNA, DNA is made up of | 4 nucleotides |
| DNA , 4 nucleotides 1 | A - Adenine |
| DNA , 4 nucleotides 2 | T - Thymine |
| DNA , 4 nucleotides 3 | C - Cytosine |
| DNA , 4 nucleotides 4 | G - Guanine |
| A = | T ALWAYS |
| C= | G ALWAYS |
| DNA Replications Major Players | DNA, DNA polymerase, Nucleotide Bases (A's, T's, C's G's) |
| DNA Polymerase...what is it? It is the enzyme in your body responsible for | naturally replicating your DNA |
| DNA Polymerase...what is it? It is the most accurate enzyme making less than | one mistake for every billion bases |
| DNA Polymerase...what is it? It has a built in | proofreading capability |
| DNA Replication, Where to start and stop? Mother nature uses | "codons" |
| DNA Replication, Where to start and stop? Scientists use | primers |
| Primers let scientists designate where they would like | amplification to begin and end. |
| What is PCR? | The amplification of a DNA sequence |
| What is in a PCR tube? | DNA, Primers, A's T's C's and G's, DNA Polymerase |
| PCR reaction, PCR is broken into | cycles |
| PCR reaction 94°C for 1 min | denaturation |
| PCR reaction 55°C for 20sec | annealing |
| PCR reaction 72°C for 2min | elongation |
| Taq Polymerase The reason that | PCR is possible |
| Taq Polymerase was Found in the bacteria that exist in | hot springs |
| PCR Applications, Why would you ever need to amplify the whole genome? | Forensics, Archeology, Disease Diagnosis |
| Who Pioneered PCR? | Dr Kary B. Mullis |
| Dr Kary B. Mullis Won the | Nobel Prize for Chemistry in 1993. |
| PCR in Research Scientists rarely have a need to look at | the whole genome |
| PCR in Research Scientists want to look at expression of | specific genes. |
| How are specific genes expressed? Does every cell carry the entire genetic code? | YES |
| How are specific genes expressed? Does every cell express the entire genetic code? | NO |
| (RT-PCR) What does the RT stand for? | Reverse Transcription |
| Reverse Transcription (RT-PCR)How? | Reverse Transcriptase |
| Reverse Transcriptase | Retro-viruses have it |
| Reverse Transcriptase VERY | sloppy enzyme, 1 mistake in every 2,000 bases |
| Inside an RT-PCR tube Ingredients | mRNA, Primers, Taq, Reverse Transcriptase, dNTP's |
| Reverse Transcriptase, Primers, and dNTP's make the first strand of | cDNA. |
| After the first strand of cDNA is made the other primer binds and Taq helps to make the first | dsDNA |
| After the creation of dsDNA, PCR continues on as before | Denaturing, Annealing, elongation |
| (Primers) It is generally accepted that if you find "part" of the message you have found | the entire message |
| (Analyzing RT-PCR)What is Gel Electrophoresis? | Forcing molecules through a gel with electric charge. |
| Analyzing RT-PCR Big/Heavy molecules vs | Small/Light molecules. |
| What charge does DNA carry? | Negative |
| How do we see DNA in a gel? | Ethidium Bromide |
| Ethidium Bromide, How does it work? | Intercalating agent.......Fluoresces under UV light |
| Analyzing a gel | DNA bp ladder, Positive Control, Negative Control, Samples |
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