Set1 is a histone methyltransferase that specifically methylates histone H3 at t
ID: 259323 • Letter: S
Question
Set1 is a histone methyltransferase that specifically methylates histone H3 at the lysyl residue located at the fourth position (H3K4) within the exposed tail. Set1 can mono-, di-, or trimethylate H3K4 and this degree of methylation can impact nucleosome positioning and thus gene expression level.
The multiple cloning site (MCS) is contained specifically in which region of this plasmid: _______________ . This is particularly useful for ________ selection process where cells could be grown on __________ media source.
b. The SET1 gene contains _________
c. The insert and pRS406 plasmid were digested with the restriction enzymes SacII and XhoI to create the homologous sticky overhangs necessary for cloning. Using the pRS406 vector map, determine the size of the linearized pRS406 plasmid produced from this double digest.
d. Ligation reactions are set up to complete the cloning of the insert into the pRS406 plasmid. What is the size of the newly designed pRS406-set1plasmid?
e. This plasmid is utilized for yeast strains, where it contains the URA3 gene that can allow for strains that are deficient in uracil biosynthesis (ura3?) to be able to biosynthesize uracil when the plasmid is taken up by the host cell. Could this plasmid also be utilized in bacteria? How can you tell?
Created with SnapGene 4319) Aat Pfol (46) (4317) Zra Ndel (330) Bo (363) (3996) Xmni sbtI (399) BfuAI BspMI (402) XcmI (545 PpuMI (570) Ncol (622) BstBl (682) Bbsl (691) BsmI (779) 3545) NmeAIII Stul (853) (3397) Ahdl pRS406 4384 bp Bpu10I (1147) BtrBI (1242) Nsil (1244) 413 rev MCS BsaAI (75) BtgzI (156) lac promoter NgoMIV (166) Nael (1678) Acc651 (2004) KpnI (2008) Sall (2025) BspDI- ClaI (203) (2040) HindIII EcoRI (2052) TspMI Xmal (2064) Smai (2066) BamHI (2070 Spel (2076) Xbal (2082) EagI- NotI (2089) Alel (2100) SacII (2101) BstXI (2102) Eco53kI (2108) Sad (2110)Explanation / Answer
a. The multiple cloning site (MCS) is contained specifically in which region of this plasmid: between 2004 to 2110 (no. mentioned in plasmid map). This is particularly useful for blue white selection process where cells could be grown on x gal and IPTG media source.
b. The SET1 gene contains 3243 nt (question is not clear to me)
c. Restriction dogetion will take 82 nt out from the vector so final size will be 4302 nt.
D. 4302+3243=7545
E. it can utilized in bacteria, selection can be based on ampicillin marker.
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