Show:
Vocabulary
Transfer of genetic information
Regulation of Gene Expression
Recombinant DNA technology
Restriction Mapping
100
What is a phenotype and genotype?
Phenotype-physical expression Genotype- genetic expression
100
How is genetic diversity introduced in cells that replicate by binary fission. (hint: 3 things)
1- genetic transfer 2- recombination of plasmids 3- transposons
100
What two genes did we focus on for exam three?
The Lac gene and the arabinose gene
100
What is PCR?
polymerase chain reaction- replicates DNA
100
Why do you avoid cutting a resistance gene with a restriction enzyme?
The bacteria would lose resistance and there would be no way of selecting colonies that incorporated the plasmid.
200
What is an operon?
"a unit made up of linked genes that is thought to regulate other genes responsible for protein synthesis."
200
Three methods of genetic transfer
1- transduction 2-conjugation 3- transformation
200
What is a global regulatory mechanism. (Hint: regulating gene expression)
Regulon- a single regulatory protein that controls many operons
200
What are the three methods that recombinant DNA is made?
1-Transformation, 2-Phage Introduction, and 3-Non-Bacterial Transformation.
200
What is the purpose of ethidium bromide?
It stains the dna to be visualized
300
What is cDNA?
complementary DNA (cDNA) is DNA synthesized from a messenger RNA (mRNA) template in a reaction catalysed by the enzymes reverse transcriptase and DNA polymerase.
300
Name at least one way DNA is introduced into the cell in transformation. (Explain how it works)
natural uptake of naked DNA electrocompetance calcium competance
300
How does glucose effect Lac gene expression?
+ glucose= +lac - glucose= - lac
300
What are the steps in genetic engineering? (hint: there are 5)
(1) DNA isolation ?(2) Restriction digestion (3) Vector insertion (4) Molecular cloning (5) DNA analysis:
300
Explain how electrophoresis works?
Based on charge and size. (DNA negative charge for example size because of pores of matrix)
400
What is a plasmid and a vector?
A plasmid is a genetic structure in a cell that can replicate independently of the chromosomes, typically a small circular DNA strand in the cytoplasm of a bacterium A vector is a DNA molecule used as a vehicle to artificially carry foreign genetic material into another cell, where it can be replicated and/or expressed
400
What is the difference between the lytic cycle and lysogenic cycle?
Lytic cycle will kill the cell lysogenic will coexist untill triggered to lytic cycle
400
What are the three major levels of gene regulation?
-autogenous regulation -repression -activation
400
What is whole gene shotgun sequencing?
Skips the mapping step of clone-based sequencing. Instead, (1) clones millions of the genome's small fragments in plasmids (2) sequences all of these small overlapping fragments, and then (3) uses computers to find matches and join them together.
400
Calculate the TM- 5' ATGGCGTAAC
2(A+T) + 4(G+C) 2(5) + 4(5) 30 degrees Celsius
500
What is sigma-32
Sigma-32 is a heat shock protein that DnaK binds to
500
What is transduction?
also known as transfection: Usage of a virus vector to deliver the DNA into the target cell.
500
Explain the difference of CAP classes.
Class 1-require only CAP for activation and is found upstream of the promoter Class 2- require only CAP for activation but overlap with promoter Class 3-require CAP and other regulators
500
What are the cycles of PCR?
1) Denature- 95 degrees for 5 mins 2) Denature- 1 min 3) anneal- TM (2(A+T)+4(G+C)) 4) extension- 72 degrees (KB/minute) 5)Extension 72 degrees for 10 minutes
500
Procedure for cloning a gene into a vector.
1-Design Primers 2-restrict 3-purify 4-ligate (PCR) 5-transform and plate 6- select 7-PCR screen 8- plasmid prep 9-sequence