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Would transfected plasmids bind histones in HeLa cells?


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#1 bsksln

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Posted 15 July 2009 - 10:21 PM

Hi all,
I am new to this field. I've read the 2004 cell paper on H3K4me1/2 demethylase LSD1 identification, and have a question on one of the experiment.


In this experiment, (figure1C, D), the author transfected G4-LSD1 and G4DBD plasmids into HeLa cells together with G4TK-Luc reporter gene. The result shows that reporter luciferase activity decreased after G4LSD1 transfection. I guess that the promoter of this reporter gene must locate in the nucleus and be bound with K4 methylated histone H3, so that LSD1 demethylase activity can act to repress the reporter gene.
I am confused about this point. How can the transfected G4TK-Luc reporter gene get into the nucleus and associate with K4 methylated histone? Or there is anthor explanation about this experiment?
Thanks!
;)

Edited by bsksln, 16 July 2009 - 01:26 AM.


#2 Dr Teeth

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Posted 16 July 2009 - 06:41 AM

Reporter genes enter the nucleus during cell division when the nuclear envelope is broken down.

Science is simply common sense at its best that is rigidly accurate in observation and merciless to fallacy in logic.
Thomas Henry Huxley

#3 bsksln

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Posted 17 July 2009 - 06:53 PM

Reporter genes enter the nucleus during cell division when the nuclear envelope is broken down.


Thanks Dr Teeth!
but will the reporter gene integrate into the genome, or will it be bound with histones?
would the G4LSD1 get into nucleus?
i just don't understand how it works.

#4 mikew

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Posted 21 July 2009 - 12:24 PM

Histones integrate into plasmid DNA in the nucleus.
But apparently the placement isn't the same as the
endogenous locus. But they're there.




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