A salivary amylase transgene is efficiently expressed in liver but not in parotid gland of transgenic mice.
Two distinct mouse amylase cDNAs, corresponding to the genes Amy-1.1 and Amy-1.2, have been isolated from a YBR/Ki parotid cDNA library. A cosmid clone containing the intact Amy-1.1 gene from strain YBR/Ki, including both the parotid and liver promoters, was transferred to the germ line of C57BL/6J mice. Two independent transgenic lines were produced. The transferred genes are organized as a 4-copy autosomal locus in line Tg518 and an 8-copy Y-linked locus in line Tg2736. Serum of both transgenic lines contained high levels of the AMY1B isozyme encoded by the transferred gene. Transcripts were detected in liver and, at a lower level, in several other tissues including white and brown fat. The anticipated expression in parotid was not observed. Constructs containing 270 or 540 bp of the 5' flanking region of the parotid promoter, cloned upstream of the chloramphenicol acetyltransferase (CAT) structural gene, were also not expressed in transgenic mice. The results suggest that sequences located more than 5 kb upstream of the Amy-1 parotid promoter and/or more than 10 kb downstream from the structural gene are required for parotid-specific expression. The results also demonstrate that non-parotid sources can produce a normal level of AMY1 in serum. Liver is the probable source of AMY1 in serum of these transgenic mice.
Jones
J M
JM
Department of Human Genetics, University of Michigan Medical School, Ann Arbor 48109-0618.
Keller
S A
SA
Samuelson
L C
LC
Osborn
L
L
Rosenberg
M P
MP
Meisler
M H
MH
D000681:Amylases / Q000235:genetics*; D000818:Animals; D015139:Blotting, Southern; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D005260:Female; D005796:Genes; D008099:Liver / Q000201:enzymology*; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D009928:Organ Specificity; D010179:Pancreas / Q000201:enzymology; D010306:Parotid Gland / Q000201:enzymology*; D010375:Pedigree; D015183:Restriction Mapping; D012463:Saliva / Q000201:enzymology*; D014158:Transcription, Genetic
D004247:DNA; D000681:Amylases