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
