The Characterization of Genomic Human Angiotensinogen Clones by Restriction Mapping
Loading...
Authors
Schadewald, Susan A.
Issue Date
1986
Type
Thesis
Language
en_US
Keywords
Alternative Title
Abstract
Human genomic angiotensinogen has been cloned into the
EcoRI site of lambda Charon 4A in the laboratory of Dr. K. R.
Marotti (The Upjohn Company) in order to study the biosynthesis
of angiotensinogen and its possible role in the pathogenesis of
human hypertension. The purpose of this study was to
characterize the four human genomic angiotensinogen clones by
restriction endonuclease cleavaqe analysis and hybridizing
experiments. The four clones have been labeled 5, 7, 13, and
14. Clones 5, 7, and 13 were determined to be identical
fragments of the angiotensinogen gene. Althouqh clone 14
includes the same fragment as the other clones, it extends
approximately 1100 nucleotides more at the 5'- end. The total
length of the angiotensinogen gene fragment of clones 5, 7, and
13 is approximately 3700 base pairs, while that of clone 14 is
approximately 4800 basepairs long.
Only the 3'- end of the human angiotensinogen cDNA, of
1571 base pairs, hybridizes with the four clones, because only
the PstI fragments of the cDNA from nucleotides 589 to 1213
(624 base pairs) and from 1213 to 1571 (358 base pairs) were
found to hybridize with the clones. It was also determined
that the restriction enzymes BamHI, BqlII, EcoRV, HindIII and
PvuI have no cleavage sites within any of the four clones. Two
different, equally plausible restriction maps for each of
clones 13 and 14 were constructed incorporating the cleavage
sites of the enzymes EcoRI, PstI, XmnI, BqlI and StuI. These
restriction maps will enable the specific fragmentation of the
clones as a primary approach to determining the sequence of the
nucleotides making up the human angiotensinogen gene.
Description
iv, 26 p.
Citation
Publisher
Kalamazoo College
License
U.S. copyright laws protect this material. Commercial use or distribution of this material is not permitted without prior written permission of the copyright holder.