Path: utzoo!utgpu!watmath!iuvax!bionet!hkucc.uucp!hrmbdkc
From: hrmbdkc@hkucc.UUCP
Newsgroups: bionet.molbio.methds-reagnts
Subject: TRANSFECTION AND CAT ASSAY
Message-ID: <8904251902.AA28874@hkucs.HKU.HK>
Date: 25 Apr 89 19:02:12 GMT
Sender: daemon@NET.BIO.NET
Lines: 24

I WANT TO TRANSFECT CAT PLASMIDS INTO RABBBIT CHONDROCYTES.
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FEATURES OF CHONDROCYTES:-
1. CHANGE PHENOTYPE (DEDIFFERENTIATE) WHEN ALLOWED TO SIT DOWN
   ON CULTURE DISH.
2. FORM AGGREGATE WHEN ALLOWED TO GROW IN SUSPENSION.

THE PREPARATION OF PRIMARY RABBIT CHONDROCYTES IS VERY TIME-
CONESUMING. IT TAKES ONE WHOLE DAY TO DISSECT 3 RABBITS AND GET
180 MILLION CELLS.

WHAT IS YOUR SUGGESTION OF CHOOSING WHICH TRANSFECTION METHOD THAT
I SHOULD USE? AND PLEASE GIVE ME HINTS HOW TO DO THAT.

CAT ASSAY USING SCINTILLATION METHOD
------------------------------------

I DOUBT THAT THE LEVEL OF EXPRESSION OF MY CAT PLASMIDS IN
THE RABBIT CHONDROCYTES NOT HIGH. BY USING WHAT MODIFICATIONS
CAN I MEASURE THIS LOW ACTIVITY.

THERE ARE T7 AND T3 PROMOTER SEQUENCES IN MY CAT PLASMIDS.
DO YOU THINK THIS WILL SEVERELY AFFECT THE ASSAY.