Construction Of Forebrain-specific RtTA Expression Transgenic Mouse And Its Preliminary Application

Regulated gene expression is an important experimental approach for researching accurately functions of particular gene products in special time of development and special tissue, at the same time, this conditional gene expression may help to avoid not only blocking analysis of phenotype caused by toxic or lethal effects in embryonic development early time, but also complications phenotype caused by ubiquitous overexpression. Tet system, from multitudinous regulative system blooming, is widely applied to each biology domain because of tightly regulated control of gene-expression. And this tet system has been applied successfully expression of extraneous gene in transgenic mouse.And then we utilized the benefits of an optimized tet-on system to construct p279-rtTA with molecular biology method. Reverse tetracycline-responsive transcriptional activator (rtTA) gene cutting from pBC-rtTA was effectively inserted into the vector of 279 containing CaM kinase IIalpha promoter to create the rtTA expressing vector, i.e., p279-rtTA. CaM kinase IIalpha -rtTA transgenic fragment (12.9 kb) released from p279-rtTA was transferred into FVB mice by pronucleus microinjection to produce transgenic mouse, to this constructs forebrain-specific rtTA transgenic tool mouse. The other type of transgenic mice containing pTRE-SV40Tag in this Tet-on system had been obtained before. The two types of transgenic mice crossed, their subsequent generation (F1) containing the two extraneous gene construction were induced by doxycycline in the diet. Through detecting the expression of SV40Tag in the F1 mice to prove the induction efficiency of this transgenic tool mice.