Zhou 3-10

Question 1

Who created the means of iPS production from adult cells? What factors were used?

Answer 1

Shinya Yamanaka Nobel prize 2012, TF’s were added Oct4, Sox2, KLF4, Myc

Question 2

Barcode language of chromosome conformation

Answer 2

the modification of the histones

Question 3

How is chromatin changed?

Answer 3

By combined action of the TF and the Chromatin modifying (editor) enzyme

Question 4

What domains make up TF’s?

Answer 4

DNA- binding domain (HLH- one helix gives specificity, one gives affinity), and activatior domain (gives ability to connect with editor proteins to form an activating or repressing complex)

Question 5

HAT

Answer 5

histone acetyl transferase- opens chromatin

Question 6

SWI/SNF

Answer 6

ATPase, uses to slide the nuclesome to allow promoter to be exposed

Question 7

Promoter

Answer 7

basal signal to start transcription binds RNA polymerase

Question 8

Proximal Promoter

Answer 8

upstream of promoter part of the promoter

Question 9

-30 site

Answer 9

basal transcriptional apparatus containing TATA box

Question 10

Activator

Answer 10

binds enhancer and mediator complex which loops to connect with the RNA Pol intiation complex

Question 11

TBP

Answer 11

Tata binding protein

Question 12

TAFs

Answer 12

TBP associated factors

Question 13

Techniques used to identify TF binding sites in vivo

Answer 13

ChIP- use sonication to break the DNA to small pieces and precipitate with Ab’s, Luciferase Reporter Assay- transfect cell with promoter sequence and luciferase gene, give luciferase substrate and see if fluoresces

Question 14

Techniques used to identify TF binding sites in vitro

Answer 14

Electrophoretic Mobility Shift Assay- reagents= TF, Ab, probe clones, competitor sequence, noncompetitor sequence, creates shift if TF binds and isupershift if the Ab has joined with the TF DNA Footprinting- uses DNAase and end-labelled P-32 DNA. creates gaps in the banding pattern on the gel where TF’s bind