aberrant migration of peptides in PAGE

Luis Branco branco at ent.umass.edu
Fri Sep 9 10:48:46 EST 1994


I have been characterizing two insect virus genes with novel regulatory
regions.  As part of this analysis, I have hybrid selected mRNAs specific to
those genes and have translated them in vitro using a rabbit reticulocyte
lysate.  The migration of both polypeptides in SDS-PAGE is difficult to
interpret, since they are falling 10 to 12 KDa below the predicted size as
determined by the ORF.  I have analyzed the DNA and amino acid sequences for
all known putative regulatory mechanisms, including signal peptidase
cleavage sites and post translational modification.  Since the polypeptides
are translated in vitro, post-translational modification is unlikely in the
rabbit ret. lysate system, except maybe for myrystylation.

Has anyone observed this phenomenon in similar circumstances, and has been
able to determine the cause for the aberrant migration of polypeptides on
PAGE?

Input in this matter would be greatly appreciated.

Luis Branco

branco at ent.umass.edu
university of massachusetts, amherst, ma. usa
413-545-5856
413-545-1578 fax
                               Luis M. Branco
                 Departments of Microbiology and Entomology
                           Morrill Science Center
                           Section IV North, Rm 2
                        University of Massachusetts
                          Amherst, MA. 01003-5720

                            Phone 413-545-5856
                            Fax   413-545-1578

                      E-Mail   branco at ent.umass.edu  



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