Facilitated dna transfer to rat submandibular gland in vivo and
grp-ca gene regulation.
O'connell, Brian C., Kelly G. Ten Hagenn, Krzysztof W. Lazowski,
Lawrence A. Tabakn, and Bruce J. Baum.
Clinical Investigations and Patient Care Branch, National Institute
of Dental Research, National Institutes of Health, Bethesda, MD
20892-1190, Departments of Dental Research and Biochemistry,
University of Rochester School of Medicine, 601 Elmwood Avenue,
Rochester, NY 14642
APStracts 2:0057G, 1995.
The internalization of DNA can be facilitated by adenovirus infection.
Using the replication-deficient adenovirus, Ad-dl312, and a plasmid
-based firefly luciferase gene as a reporter, we have optimized the
uptake and expression of DNA in rat submandibular glands in vivo.
Luciferase expression is transient and peaked at 18 hours after
infection. Luciferase activity increased with plasmid concentration
and was greatest at 109 to 1010 plaque forming units of Ad-dl312 per
gland. We next examined the expression in vivo of plasmids containing
deletions of the glutamine/glutamic acid-rich protein (GRP-Ca
isoform) gene upstream region linked to a chloramphenicol
acetyltransferase (CAT) reporter. Constructs with 9.4 kilobases (kb),
6.3 kb, 2.7 kb and 17 base pairs of upstream sequence gave relative
CAT activities of 100, 30, 7.6 and 38.5, respectively. Using the 9.4
kb GRP-Ca construct, CAT was preferentially expressed in acinar
cells, which is characteristic of GRP. This gene transfer approach
should prove useful in the further study of gene expression in
salivary glands and other organs.
Received 23 December 1994; accepted in final form 23 March 1995.
APS Manuscript Number G497-4.
Article publication pending Am. J. Physiol. (Gastrointest. Liver
Physiology).
ISSN 1080-4757 Copyright 1995 The American Physiological Society.
Published in APStracts on 4 April 1995.