1. Quiz#8 LC710 10/20/10 name___________
Q1 (3pts) :I have the UBX homeodomain sequence and want to determine (in vivo) the optimal
binding sites. I know that TAATTG is a good in vitro binding site and know that TAAT is absolutely
reguired for UBX binding. Using the two plasmids below (you may modify them at will)
and a third one that you need to make, please design a clever experiment to look for the best binding sites.
Please add modifications to plasmids
is TATA minimal promoter #1
Red F.P. #2
Green F.P.
CMV is Eukaryotic promoter
CMVp #3
MCS
Please write it in the form of an outline

2. Q2:
(2pt) Below is the UBX DNA binding portion of the protein. You want to fuse it VenusFP. :
UBX:
Nt-RRRGRQTYTRYQTLELEKEFHTNHYLTRRRRIEMAHALCLTERQIKIWFQNRRMKLKKEIQ* -Ct
Genetic Code
Phe-TTC
Leu-CTG
Ile-ATC
Met-ATG
Val-GTG
Ser-TCT
Pro-CCC
Thr-ACT
Ala-GCC
Tyr-TAT
His-CAT
Gln-CAA
Asn-AAT
Lys-AAA
Asp-GAT
Glu-GAA
Cys-TCT
Trp-TGG
Arg-CGA
Gly-GGT
*(stop)-TAA
VenusFP:
Nt-MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKLICT
TGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIF
FKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHN
VYITADKQKNGIKANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNH
YLSYQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK*-Ct
Write out the -9 to +9 nucleotides around the junction
5’ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ 3’
Q3:
(2pt) You want to mutate Phe3 to Tyr3 in VenusFP. Briefly List two
Different techniques to achieve this mutation. Which is faster A or B? A) B)