EXPERIMENTS TO BE PERFORMED BY B. ROTMAN IN DR. BRITTON CHANCE 'S LABORATORY
AT PHILADELPHIA

Purpose:

 

To test the hypothesis that the reduced beta-galacto@idase activity of
intact cells of EB, Coli i@ due to inhibition of the enzyme by the reaction products,
The hypothesis predicts theéthe enzymatic activity of the intact cells will be
larger when measured at initial time because of the absence of reaction products.

Method:

About 10 liters of cells of W1317 will be grown in mineral medium, For
handling purposes the culture will be centrifuged down and will be resuspended
in a few hundred mil of the same medium. Such suspension will be kept at low
temperatures and will be carried as such to Philadelphia, Before the experiment the
suspension will be washed with buffer and will be resuspenied with buffer. The
suspension will be then mixed with OMPG in a rapid flow-double beam apparatus and
the appearance of the yellow color will be recorded after one millisecond or less.
As a control a benzene treated culture will be run in the same fashion. The blank
in each case will be cells treated or untreated mixed with buffer without ONPG.

Madison, October 26, 1955