PMID- 10854782
OWN - NLM
STAT- MEDLINE
DCOM- 20000724
LR  - 20190707
IS  - 0378-1119 (Print)
IS  - 0378-1119 (Linking)
VI  - 250
IP  - 1-2
DP  - 2000 May 30
TI  - Identification of a 3.2 kb 5'-flanking region of the murine keratocan gene that 
      directs beta-galactosidase expression in the adult corneal stroma of transgenic 
      mice.
PG  - 85-96
AB  - The mouse keratocan gene (Ktcn) expression tracks the corneal morphogenesis 
      during eye development and becomes restricted to keratocytes of the adult, 
      implicating a cornea-specific gene regulation of the mouse Ktcn [J. Biol. Chem., 
      273 (1998) 22584-22588]. To examine the functionality of the mouse Ktcn promoter, 
      we have cloned and sequenced a 3.2kb genomic DNA fragment 5' of the mouse Ktcn 
      gene, which was used to prepare a reporter gene construct that contained the 
      3.2kb 5' flanking sequence, exon 1 and 0.4kb of intron 1 of Ktcn, and beta-geo 
      hybrid reporter gene. The beta-galactosidase (betaGal) activity was assayed in 
      tissues of two of five transgenic mouse lines obtained via microinjection. In 
      adult transgenic mice, betaGal activity was detected only in cornea, not in other 
      tissues (e.g. lens, retina, sclera, lung, heart, liver, diaphragm, kidney, and 
      brain). During ocular development, the spatial-temporal expression patterns of 
      the betaGal recapitulated that of endogenous Ktcn in transgenic mice. Using XGal 
      staining, strong betaGal activity was first detected in periocular tissues of 
      E13.5 embryos, and restricted to corneal keratocytes at E14.5 and thereafter. 
      Interestingly, in addition to cornea, betaGal activity was transiently found in 
      some non-ocular tissues, i.e. ears, snout, and limbs of embryos of E13.5 and 
      E14.5 but was no longer detected in those tissues of E16.5 embryos. The transient 
      expression of endogenous keratocan in non-ocular tissues during embryonic 
      development was confirmed by in situ hybridization. Taken together, our results 
      suggest that the 3.2kb Ktcn promoter contains sufficient cis-regulatory elements 
      to drive heterologous minigene expression in cells expressing keratocan. The 
      identification of keratocyte-specific expression of betaGal reporter gene in the 
      adult transgenic mice is an important first step in characterizing the Ktcn 
      promoter in order to use it to drive a foreign gene expression in corneal stroma.
FAU - Liu, C
AU  - Liu C
AD  - Department of Ophthalmology, College of Medicine, University of Cincinnati, OH 
      45267-0527, USA. liucg@email.uc.edu
FAU - Arar, H
AU  - Arar H
FAU - Kao, C
AU  - Kao C
FAU - Kao, W W
AU  - Kao WW
LA  - eng
SI  - GENBANK/AF186468
GR  - EY 11845/EY/NEI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - Netherlands
TA  - Gene
JT  - Gene
JID - 7706761
RN  - 0 (Kera protein, mouse)
RN  - 0 (Proteoglycans)
RN  - 0 (RNA, Messenger)
RN  - 0 (Recombinant Fusion Proteins)
RN  - EC 3.2.1.23 (beta-Galactosidase)
SB  - IM
MH  - Animals
MH  - Base Sequence
MH  - Cells, Cultured
MH  - Cornea/cytology/metabolism
MH  - Corneal Stroma/*metabolism
MH  - Embryo, Mammalian/metabolism
MH  - Gene Expression
MH  - Gene Expression Regulation, Developmental
MH  - In Situ Hybridization
MH  - Mice
MH  - Mice, Transgenic
MH  - Molecular Sequence Data
MH  - *Promoter Regions, Genetic
MH  - Proteoglycans/*genetics
MH  - RNA, Messenger/genetics/metabolism
MH  - Recombinant Fusion Proteins/genetics/metabolism
MH  - Sequence Homology, Nucleic Acid
MH  - Tissue Distribution
MH  - Transgenes/genetics
MH  - beta-Galactosidase/*genetics/metabolism
EDAT- 2000/06/16 00:00
MHDA- 2000/06/16 00:01
CRDT- 2000/06/16 00:00
PHST- 2000/06/16 00:00 [pubmed]
PHST- 2000/06/16 00:01 [medline]
PHST- 2000/06/16 00:00 [entrez]
AID - S0378111900001657 [pii]
AID - 10.1016/s0378-1119(00)00165-7 [doi]
PST - ppublish
SO  - Gene. 2000 May 30;250(1-2):85-96. doi: 10.1016/s0378-1119(00)00165-7.