User:Behzad Damadzadeh/Notebook/PcTF Subcloning in E-coli/2013/08/09

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(08/09/2013)
 
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{| {{table}} cellspacing=''7''
{| {{table}} cellspacing=''7''
|- valign="top"  
|- valign="top"  
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| '''BioBrick Part''' || '''Vector''' || '''Description''' || '''Microscopy Image'''
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| '''BioBrick Part''' || '''Vector''' || '''Description''' || '''Microscopy Image''' || '''Comments"""
|-
|-
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| KAH59 || PcVN || [K : Gal4 DB : mCh : mCh] + VP64-NLS || [[Image:Merge-KAH59-PcVN.png |250px]]
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| KAH59 || PcVN || [K : Gal4 DB : mCh : mCh] + VP64-NLS || [[Image:Merge-KAH59-PcVN.png |250px]] || RFP and CFP signal
|-
|-
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| KAH60 || PcVN || [K : Gal4 DB : mCh] + VP64-NLS || [[Image:Merge-KAH60-PcVN.png |250px]]
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| KAH60 || PcVN || [K : Gal4 DB : mCh] + VP64-NLS || [[Image:Merge-KAH60-PcVN.png |250px]] || RFP and CFP signal
|-
|-
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| KAH54 || PcVN || [[K : Gal4 DB] + VP64-NLS || [[Image:KAH54-PcVN.png |250px]]
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| KAH54 || PcVN || [[K : Gal4 DB] + VP64-NLS || [[Image:KAH54-PcVN.png |250px]] || CFP signal only
|-
|-
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| KAH59 || PcN (MV5) || [K : Gal4 DB : mCh : mCh] + NLS || [[Image:Merged-KAH59(MV5).png |250px]]
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| KAH59 || PcN (MV5) || [K : Gal4 DB : mCh : mCh] + NLS || [[Image:Merged-KAH59(MV5).png |250px]] || RFP signal only
|-
|-
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| KAH66 || PcN (MV5) || [K : Gal4 DB : mCh : mCh : VP64] + NLS || [[Image:Merged-KAH66-MV5.png |250px]]
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| KAH66 || PcN (MV5) || [K : Gal4 DB : mCh : mCh : VP64] + NLS || [[Image:Merged-KAH66-MV5.png |250px]] || RFP and CFP signal
|}
|}
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* Since RFP and CFP expressed at the same time, I concluded that both plasmids transfected to the U2OS cells and VP64 activated CFP expression.
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KAH54 has no mCherry sequence but VP64 could activate CFP expression and KAH59(MV5) without VP64 only shows RFP signal.

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08/09/2013

Flourescent Microscopy

  1. Check RFP/CFP of co-transfection after 18 hrs.


BioBrick Part Vector Description Microscopy Image Comments"""
KAH59 PcVN [K : Gal4 DB : mCh : mCh] + VP64-NLS RFP and CFP signal
KAH60 PcVN [K : Gal4 DB : mCh] + VP64-NLS RFP and CFP signal
KAH54 PcVN [[K : Gal4 DB] + VP64-NLS CFP signal only
KAH59 PcN (MV5) [K : Gal4 DB : mCh : mCh] + NLS RFP signal only
KAH66 PcN (MV5) [K : Gal4 DB : mCh : mCh : VP64] + NLS RFP and CFP signal
  • Since RFP and CFP expressed at the same time, I concluded that both plasmids transfected to the U2OS cells and VP64 activated CFP expression.

KAH54 has no mCherry sequence but VP64 could activate CFP expression and KAH59(MV5) without VP64 only shows RFP signal.

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