Term
| Who founded Genentech? What did he discover? |
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Definition
| Herbert Boyer. Discovered EcoRI. |
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Term
| Recognition sequence for EcoRI |
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Definition
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Term
| What does RFLP stand for? |
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Definition
| Restriction-Fragment Length Polymorphism |
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Term
| HindII recognition sequence |
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Definition
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Term
| EcoRV recognition sequence |
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Definition
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Term
| What are the cofactors needed for Type I, Type II, Type III? |
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Definition
Type I: SAM, ATP, Mg2+
Type II: Mg2+
Type III: Mg2+ |
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Term
| What people won Nobel Prizes? For what? |
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Definition
Werner Arber - Type I
Daniel Nathans/Hamilton Smith - Type II
Richard Roberts - split genes
Paul Berg - recombinant DNA
Michael Smith - site-directed mutagenesis
Roger Kornberg - eukaryotic transcription
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Term
| PvuII recognition sequence |
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Definition
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Term
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Definition
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Term
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Definition
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Term
| Where do the following cleave: Type I, Type II Type III? |
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Definition
Type I: 1000 bp away
Type II: at site
Type III: 24-26 bp away |
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Term
| Name examples of the following cloning vectors: 1st, 2nd, 3rd, 4th generation. |
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Definition
1st: pSC101
2nd: pBR322
3rd: pUC8, 9, 18, 19
4th: pUC118, 119, pBluescript, pTZ18, 19 |
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Term
| What is the name of the lambda repressor? |
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Definition
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Term
| Which promotes the lytic cycle, cro or cI? |
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Definition
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Term
| Out of the promoters Pr and Prm, which promotes cro and which promotes cI? |
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Definition
Pr promotes cro.
Prm promotes cI. |
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Term
| What does "YAC" stand for? Why is is important. |
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Definition
| Yeast Artificial Chromosome. Can enable expression of larger eukaryotic genomes (better glycosylation). |
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Term
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Definition
| Discovered cancer transcription patterns different than normal cells. |
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Term
| What does "PD...D/ExK" mean? Which residue is most important? |
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Definition
| Proline, Asp...Asp/Glu, anything, Lysine |
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Term
| What did Berg get Nobel for? |
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Definition
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Term
| Name viruses of the following types: dsDNA, ssDNA, dsRNA, and 3 types of ssRNA |
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Definition
dsDNA: Herpes, Pox, Adeno, SV40
ssDNA: Roseola
dsRNA: Reo
ssRNA (acts as RNA): Polio, Rhinovirus
ssRNA (template for mRNA):Rabies, Measles, Flu
ssRNA (template for DNA): HIV |
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Term
| What do N and Q do in lambda phage? |
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Definition
| Produce anti-terminator proteins. |
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Term
| What is the major repressor of the lambda lytic cycle? |
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Definition
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Term
| What is the name of the repressor of the lambda repressor? |
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Definition
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Term
| What are the steps of induction in lambda phage? |
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Definition
1. Damage to DNA (UV, etc.)
2. RecA protease attacks cI
3. Cro comes in
4. Lysing occurs. |
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Term
| What are the two ways viruses can change their antibodies? |
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Definition
1. Antigenic shift: The antibody gene is replaced entirly by a new gene.
2. Antigenic drift: Point mutations occur naturally. |
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Term
| How do you use homo-polymer tailing to join to blunt-end DNA strand? |
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Definition
| Terminal transferase adds homo-polymer tails to both pieces of DNA. This makes sticky ends. Use ligase to stick them together. |
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Term
| What does dpnI do in the PCR process? |
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Definition
| Eats away methylated strands at end of rxn. |
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Term
| Southern Blotting tests for... |
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Definition
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Term
| Northern Blotting tests for... |
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Definition
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Term
| Western Blotting tests for... |
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Definition
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Term
| Draw and properly name X-Gal. |
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Definition
| 5-bromo-4-chloro-3-indoyl-B-D-galactose |
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Term
| What are the general features of pUC18? |
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Definition
| lac gene with MCS (multiple cloning site), amp (anti-biotic resistasnce), ori |
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Term
| What are the general features of pUC118? |
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Definition
| lac gene with MCS (multiple cloning site), amp (anti-biotic resistasnce), ori, M13ori |
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Term
| What do DNA gyrase and helicase do? Which of the two needs ATP? |
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Definition
Helicase comes in and "jacks open" the DNA. Gyrase work on the back end, cutting the double strand DNA to relase supercoling (makes linking number -2).
Gyrase needs ATP. |
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Term
| What is PRPP? What is it for? |
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Definition
| 5-phosphoribosylpyrophosphate (a ribose with 2 P's on one end, 1 P on the other). It transfers phosphate groups in the synthesis of AMP, GMP, IMP, OMP, UMP, etc. First committed step in PURINE biosynthesis. |
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Term
| What is IMP? What is it for? |
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Definition
| Inosinse monophosphate. Intermediate in PURINE biosynthesis. Converted to AMP or GMP in "branch point" of purin synthesis. |
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Term
| What is OMP? What is it for? |
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Definition
| Ornithine monophosphate. Intermediate in PYRIMIDINE biosynthesis. |
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Term
| What are the purines? What are the pyrimidines? (Remember mnemonic!) |
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Definition
puRines: A, G (RAG)
pYrimidines: U, C (YUC) |
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Term
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Definition
| Build up of uric acid. Excess PRPP causes excess nucleotide synthesis, which causes excess uric acid. |
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Term
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Definition
| Allopurinal. It stops hypoxanthine from being converted to uric acid (acts as analog inhibitor). |
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Term
| How was lac discovered? By who? |
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Definition
| Specialized transduction. Shapiro. |
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Term
| Which mutagens cannot mutate pure DNA? |
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Definition
| The base analogs (need replication) and nitrosoguanidine (needs cellular machinery to work). |
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Term
| What mutagens can work on pure DNA? |
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Definition
| EMS (Ethylmethanesulfinate), Aflatoxin, etc. |
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Term
| How does Eckstein mutagenesis work? |
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Definition
| You use a mutagenic primer and replicat in special "heavy" dCTPas. Then you use a "pac-man" to chew away the non-heavy strand, getting rid of non-mutated DNA. Exo3. |
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Term
| How does Kunkel mutagenesis work? |
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Definition
| Use M13 phage to get your start DNA. Add mutagenic primer. Grow in "-duT, -ung" E. Coli. duT represses something that would put U's everywhere. ung would normally cut out U's. But since the E. Coli is "minus" for these, the U's get put everywhere there is supposed to be a T. |
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Term
| How does Quik-Change work? |
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Definition
| Use double-stranded plasmid and forward and reverse mutagenic primers. |
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Term
| What gene is put in GMO corn and cotton. Where is is form? |
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Definition
| "Cry" gene from BT bacteria = protects from insects. |
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Term
| Who made S&M (site-directed mutagenesis)? |
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Definition
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Term
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Definition
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Term
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Definition
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Term
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Definition
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Term
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Definition
| ssRNA (acts as its own mRNA). Polio, Rhino. |
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Term
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Definition
| ssRNA (is template for mRNA). Flu, Measles, Mumps. |
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Term
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Definition
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Term
| What type of virus is Flu? |
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Definition
| Orthomyxo. Class 5 (ssRNA is a template for mRNA). |
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Term
| What on the flu virus is recognized by the human immune system (i.e. what are the antigens)? |
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Definition
| HA and NA. These mutate frequently, which is why there are different flus every year. |
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Term
| What are the general features of a tRNA? |
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Definition
| Top = acceptor loop (attaches to amino acid). Left = D-loop. Bottom = anti-codon loop. Right = T-psi-C loop. |
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Term
| What is an inactive X chromosome called? |
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Definition
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Term
| What is "dosage compensation"? |
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Definition
| To regulate genes on the two sets of female X chromosomes, one of the chromosomes is inactivated. |
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Term
| How is differentiation of cells accomplished in eukaryotes? |
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Definition
| By regulating transcription of certain genes. |
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Term
| What is the sequence of elements in the Class II core promoter of eukaryotes? |
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Definition
| TFIIB recognition, TATA box, Initiator, Down-stream core promoter |
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Term
| What kind of eukaryotic genes would you expect to lack TATA boxes? |
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Definition
| "Housekeeping genes" that are always on. I.e. they don't need to be turned on/off. |
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Term
| What are the sequence of steps of transcription of a gene with introns? |
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Definition
| 1. Transcribe entire gene. 2. Put on Cap and Poly-A Tail. 3. Splice out introns. |
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