A Novel
Bifunctional in vivo Mutation Detection System
YAO Zhen-Zhen, LI Huai-Xing,
CHE Wen-Liang, HE Yan, FU Ji-Liang*
( Department of Medical Genetics, Second Military Medical
University, Shanghai 200433,
China )
Abstract A shuttle
plasmid pMCLacI/neo with two copies of LacI was integrated into
mouse genome and a novel system which could detect in vivo mutation of
both expression and silence genes was constructed, enabling the comparative
analysis of their mutation spectra and mutant frequencies. 486 fertilized eggs
from C57BL/6 mice with microinjected pMCLacI/neo plasmid were
transferred into oviducts of 18 pseudo pregnant mice, and 32 alive offsprings
were screened and identified by using PCR and Southern blotting. Genomes of 5
mice had pMCLacI/neo plasmid integrated, as verified by Southern blot
after the PCR screening. Only one of the two LacI in pMCLacI/neo
was in expression state; and this established a model, that the status in
vivo of both gene expression and silencing could be simulated. This kind of
mice might be used as a novel bifunctional mutation detection system in vivo.
Key words transgenic mouse; mutation detection; LacI;
plasmid
*Corresponding author:
Tel, 86-21-25070027; Fax,
86-21-25070027; e-mail, [email protected]