1) You are doing FRET and you excite a protein tagged with blue fluorescent prot
ID: 272188 • Letter: 1
Question
1) You are doing FRET and you excite a protein tagged with blue fluorescent protein with 360 nm light and it emits ight with a wavelength of 450 nm. which of the following fluorescent proteins (excitation nm/emission nm) could you use to detect a near protein? (a) Red fluorescent protein (500 nm/600 nm) (b) Yellow fluorescent protein (520 nm/580nm) (e) Green fluorescent protein (450nm/700 nm) (d) Orange fuorescent protein (360 nm/450 nm) difference in a proteins function? (a) E->A 2) You are performing alanine-scanning mutagenesis. Which mutation would you least expect to make a large (b) K->A (c) P->A 3) Which of the following is common purpose for a domain swapping study? (a) testing functional equivalencies (b) testing for DNA binding (c) testing for promoter elements (d) testing for protein expression 4) Why is the restriction endonuclease Dpnl used for site-directed mutagenesis? (a) it digests palindromic sequences (c) it digests methylated DNA (b) it digests very rare DNA sequences (d) its recognition sequence is found in many MCS's. 5) Semi-quantative PCR (or RT-PCR) is normally done for how many PCR cycles? (a) 10-15 6) Which type of DNA footprinting can reveal the actual nucleotides that a protein makes contact with? (b) 15-20 (c) 20-25 (d) 25-30 (a) DNAse (b) EcoRI (c) hydroxyl radical (d) S1 nuclease 7n) Which of the following reporter genes would be the least Ikely used for a transcription assay? (b) chloramphenicol acetyl-transferase 8) Which of the following techniques might you use to identify novel protein-RNA interactions? (a) B-galactosidase (c) histidine (d) luciferase (a) ChIP (b) RIP (c) RNA footprinting (d) Yeast one-hybrid 9) Which of the following would not be studied using transcription assays? (a) promoter sequences (b) transcription factors (d) steps involved in activating a gene (c) luciferase activity 10) Which type of detection method is the "best for qPCR or qRT-PCR? (a) ethidium bromide (b) fluorescent probes (c) sybr green(d) TaqmanExplanation / Answer
1. The answer is c.) Green flouroscent protein, as FRET always detect light that is in visible range.
2. The answer is d.) I to A. That is Isoleucine to Alanine, beacuse Isoleucine is also an apolar amino acid ike alanine, though is the given options Proline is also non polar, but it carries a aromatic group.
3. The answer is d.) testing for protein expression as domain swapping is the study if protein oligomerization and some proteins become functional/non sunctional only in oligomer conditon.
4. c.) It digests methylated DNA. In E.Coli it specifically digests sequence GATC, only when A is methylated.
5. d.) 25-30 PCR cycles.
7. The answer is c.) histidine, it is an essential AA not synthesized by the body.
9. The answer is b.) transcrption factors.
10. The answer is c.) Sybr green
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