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(This message from Seima Itami in Osaka Univ.
I posted message instead of him.
Takeshi Omasa, Dr. , Dept.of Botech. Osaka univ. JAPAN)
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I'm Seima Itami, and I'm a graduated student
in Osaka University, JAPAN.
I would like to ask for the flow cytometry
for CHO (Chinese Hamster Ovary)
It's well known that the DHFR gene in the CHO cells
increases under high MTX (which is inhibitor of dhfr protein)
concentration culture.
(It's called the gene amplification).
There are two types of the amplified gene in CHO cell.
One is recombination into the complete chromosome region
(it is called homogeneously staining region; HSR),
and the other is into the extra chromosomal element
(called double minute; DM).
The length of the chromosome contains HSR is about 100 Mb,
and the extra chromosomal element is about 1 Mb.
And there are 10 to 100 copies of DHFR gene on the two regions.
I would like to classify the two types of gene ( HSR and DM),
and to quantify the copy numbers on the two regions
using flow cytometry.
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If you have some information about this, would you please teach me!
(The flow cytometer which we used was coulter EPICS elite.
This machine have both air-cooled and water-cooled laser.)
Thank you for your cooperation.
Seima Itami, Depatment of Biotech. Osaka University
e-mail itami.bce@bce.bio.eng.osaka-u.ac.jp
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