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Strain
C57BL/6-Tg(pBSKS-Slc10a2/OVA)

Summary Data Summary
Official Name C57BL/6-Tg(pBSKS-Slc10a2/OVA)
Common Name OVA-BIL
Description Slc10a2 drives expression of OVA in biliary epithelial cells

Apical sodium-dependent bile acid transporter
(ASBT)-membrane-bound ovalbumin (mOVA) complementary DNA
(cDNA) was constructed using the rat ASBT promoter,7 a
membrane- bound ovalbumin (OVA) (amino acids 139–385)8 and a
polyA tail (pDo15-polyA). The membrane-bound form of OVA
consists of a fusion protein made up of the first 118
residues of the human transferrin receptor (including
cytoplasmic tail and signal/anchor domain) linked to
residues 139–385 of mature OVA, transferrin receptor-ova
(TFR-OVA), targeting membrane expression of OVA on biliary
epithelium. The 5' flanking region of the ASBT gene was cut
using BamHI and BstXI sites (all restriction enzymes from
New England Biolabs, Ipswich, MA), resulting in a 3.0-kb
fragment that was used in the ASBT-mOVA construct. The
TFR-OVA fragment was cut from the rat insulin promoter-mOVA
plasmid8 by digestion with HindIII and XbaI, and then the
HindIII site was blunted. The ASBT promoter and TFR-OVA
fragments were then ligated into the pBSKS vector
(Stratagene, La Jolla, CA). The polyA region was a rabbit
ß-globin gene isolated from the pDOI-5 plasmid from BamHI to
XhoI and was blunted at 2 ends8 and was then inserted into
the NotI site of the pBSKS-ASBT-mOVA. Its orientation was
checked after insertion. After isolation of the plasmid,
restriction enzyme mapping was performed, and TFR-OVA was
sequenced to verify that the construct was correct. Vector
sequence was excised by digesting with BamHI and SacII. The
purified DNA was microinjected into the pronuclei of the
C57BL/6 oocytes (Jackson Laboratory, Bar Harbor, ME).
Development Status Phenotyping ongoing
Creation Method transgenic
TypeCount
Investigators 1
Experiments 1


Investigators
NameInstitution
David AssisYale University


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