cells in the middle do not express the protein

carlos h. pedemonte pedemonte at Jetson.UH.EDU
Wed Jan 26 18:53:39 EST 1994

Dear netters:
       I have a question in reference to some experiments of protein
expression we are doing. We are trying to express a protein in CV-1 cells
using the vaccinia virus/T7 RNA polymerare system. To find the the best
conditions of expression we are using a plasmid that express
b-galactosidase. We have observed that cells that are in the border of the
plates express a lot more b-gal than cells in the middle of the plate. Some
of the cells in the middle of the plate do not express the protein at all.
We think this is not due to the vaccinia or liposome solutions forming a
meniscus because we have used a good amount of solution to avoid this kind
of problem. Does anybody has any idea about this? 
Thank you, 

Carlos H. Pedemonte                     Telephone:  713-743-1211 (office)
Dept. of Pharmacology                               713-743-1228 (lab)
University of Houston                   Fax:        713-743-1229           
Houston, TX 77204-5515                  e-mail: pedemonte at jetson.uh.edu

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