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Abstract for Canine Pancreatic Microsomal Membranes
Microsomal vesicles are used to study cotranslational and initial
post-translational
processing of proteins. Processing events such as signal peptide cleavage,
membrane insertion, translocation and core glycosylation can be examined by the
translation of the appropriate mRNA in vitro in the presence of these microsomal
membranes. In addition, processing and glycosylation events may be studied by
transcription/translation of the appropriate DNA in the TNT®
Lysate Systems when used with microsomal membranes. To assure consistent
performance with minimal translational inhibition and background, microsomes
have been isolated free from contaminating membrane fractions and stripped of
endogenous membrane-bound ribosomes and mRNA. Membrane preparations are assayed
for both signal peptidase and core glycosylation activities using two different
control mRNAs. The two control mRNAs supplied with this system are the precursor
for beta-lactamase (or ampicillin resistance gene product) from E. coli and
the precursor for alpha-mating factor (or alpha-factor gene product) from S.
cerevisiae.
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