Yan ke xue bao = Eye science · 1995
mRNA expression of vimentin gene in lens of transgenic mouse and DNA amplification in human cataracts.
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Li Y, Hui Y, Zhou J, Liu X, Chen S, Hu R, Ying Z
Аннотация
PURPOSE: To investigate the role of vimentin gene in cataractogenesis.
METHODS: The 12.7kb chicken vimentin genes were microinjected into the male pronuclei of 918 fertilized mice eggs. 841 injected embryos were transferred into oviducts of pseudopregnant recipient females, of which 12 pregnant mice gave birth to 49 offspring mice. The integration and expression of exogenous gene in the offspring were analysed by Southern and Northern blot hybridizations. In the human senile cataract, the lens vimentin gene was analyzed with the chicken vimentin gene probe.
RESULTS: It showed that four of F1 offspring were transgenic mice in which the chicken vimentin gene was integrated in their genomes. The transgenic band was 12kb, similar to the 12.7kb chicken vimentin fragment injected. One 2kb vimentin mRNA was visualized on E2 mouse lens blot, which revealed that the chicken vimentin gene was efficiently expressed in this transgenic mouse. In the human senile cataract lens, 12kb BamHI-restricted vimentin fragments displayed a stronger hybridization signal than that of the control lens in Southern blot analysis. It implies that the formation of human senile cataract may be associated with the amplification of vimentin gene.
CONCLUSIONS: We have successfully developed four transgenic mice bearing chicken vimentin gene and having mRNA expression which can be used for further study. It is to be observed if the normal lens cell function is affected by the expressed product and cataract occurs in our transgenic mice. The cause of the gene amplification in human cataract remains for further investigation.
METHODS: The 12.7kb chicken vimentin genes were microinjected into the male pronuclei of 918 fertilized mice eggs. 841 injected embryos were transferred into oviducts of pseudopregnant recipient females, of which 12 pregnant mice gave birth to 49 offspring mice. The integration and expression of exogenous gene in the offspring were analysed by Southern and Northern blot hybridizations. In the human senile cataract, the lens vimentin gene was analyzed with the chicken vimentin gene probe.
RESULTS: It showed that four of F1 offspring were transgenic mice in which the chicken vimentin gene was integrated in their genomes. The transgenic band was 12kb, similar to the 12.7kb chicken vimentin fragment injected. One 2kb vimentin mRNA was visualized on E2 mouse lens blot, which revealed that the chicken vimentin gene was efficiently expressed in this transgenic mouse. In the human senile cataract lens, 12kb BamHI-restricted vimentin fragments displayed a stronger hybridization signal than that of the control lens in Southern blot analysis. It implies that the formation of human senile cataract may be associated with the amplification of vimentin gene.
CONCLUSIONS: We have successfully developed four transgenic mice bearing chicken vimentin gene and having mRNA expression which can be used for further study. It is to be observed if the normal lens cell function is affected by the expressed product and cataract occurs in our transgenic mice. The cause of the gene amplification in human cataract remains for further investigation.