[Construction of CgA gene antisense transgenic mice].
In order to get CgA gene antisense DNA transgenic mouse, we constructed the CgA gene antisense DNA plasmid pCAS2C and microinjected it into the female pronucleus of fertilized mouse eggs, and transplanted them into oviduct of the foster. Every offspring of the fosters was determined by the PCR method. The positive mice had a 300 bp DNA electrophoresis band. We selected two male positive mice from 50 offspring survived of the pseudomother. Then, two positive mice crossed with normal mice respectively to reproduce offspring of F1. All offspring of F1 were determined by PCR to select positive offspring. Positive offspring of F1 carried only one allele of pCAS2C (heterozygous pCAS2C/-). Positive F1 offspring were selfcrossed, 1/4 offspring of F2 carrying two unites of one allele pCAS2C (pCAS2C/pCAS2C) are homozygous. Then, all offspring of homozygous F2 crossed with normal mice, could produce 300 bp DNA electrophoresis band by PCR. Total RNA of brain tissue of transgenic mouse was used to RT-PCR method, the 300 bp DNA product was obtained. The result indicates that the reading frame of CgA antisense DNA of pCAS2C has expressed in the transgenic mice.
Liu
X D
XD
College of Biology, Peking University, Beijing 100871, China.
Sun
Z Q
ZQ
Zhang
C B
CB
Wu
H L
HL
D000818:Animals; D053379:Chromogranin A; D002864:Chromogranins / Q000235:genetics*; D016373:DNA, Antisense / Q000096:biosynthesis*; D018076:DNA, Complementary / Q000235:genetics*; D005260:Female; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D016133:Polymerase Chain Reaction
D053379:Chromogranin A; D002864:Chromogranins; D016373:DNA, Antisense; D018076:DNA, Complementary