應用條件化基因剔除及轉基因策略探討小鼠C型血小板生長因子(PDGF-C)生理作用之分子調控機制(2/3)
Date Issued
2004
Date
2004
Author(s)
鄭登貴
DOI
922313B002138
Abstract
Several inducible gene expression
systems have been developed in vitro in recent
years to overcome limitations with traditional
transgenic mice. One of these is the
tetracycline-regulated system that has been
used successfully in vivo, of which the
expression of the targeted protein has been
regulated by the promoter sequences containing
tetracycline responsive element. To meet these
purposes, two transgenes were first constructed,
including the former ones carries a highly
specific transcription factor (Tet- responsive
promoter) and the latter ones carries both of the
target gene of PDGF-C regulated by a Tetresponsive
promoter and the other target
equipped with a double genes containing both
the PDGF-C as well asthe EGFP reporter
sequences. Based on these, transgenic (Tg)
mice harboring the transgene of PDGF-C
assigned to be driven by tetracycline were
generated after the microinjection strategy
using a tetracycline-controlled transactivator
(rtTA) and a rtTA-dependent promoter linked
to a PDGF-C gene. These allow the expression
of PDGF-C be well regulated by the
tetracycline and/or its analogue. Consequently,
the tightly regulated PDGF-C expression can
reveal the true physiological effect of the gene.
Results from initial experiments showed
that, by double transgenic vectors co-injection
technology (Tet-on together with the transgene
of pBi-EGFP-PDGF-C and Tet-on together
with the transgene of pTRE2hyg-PDGF-C),
followed by verification with PCR technology,
in the first group, one of the twenty five new
born transgenic mouse contains both transgenes
of the Tet-on together with the
pBi-EGFP-PDGF-C. On the other hand, results
from the later studies appeared that seven out
of 65 new born mice were transgenic
containing both transgenes of tet-on and
pTRE2hyg-PDGF-C. All these transgenic mice
have reached sexually mature of age and ready
providing for further in vivo studies to elucidate
the physiological significance of PDGF-C gene
expression on events related to animal
reproduction as well as the vasculogenesis
involved in embryonic implantation.
Subjects
PDGF-C
tetracycline-regulated system
green fluorescent protein
transgenic mice
Publisher
臺北市：國立臺灣大學動物科學技術學系暨研究所
Type
report
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