波形纤维蛋白基因在转基因小鼠晶体的mRNA表达及人白内障晶体的DNA扩增

mRNA expression of vimentin gene in lens of transgenic mouse and DNA amplification in human cataract

为探讨波形纤维蛋白（ｖｉｍｅｎｔｉｎ）基因在白内障发生中的作用，用显微注射法将１２．７Ｋｂ鸡ｖｉｍｅｎｔｉｎ基因注入５３０枚小鼠受精卵的雄原核内，并将其中４５３枚卵移入１８只假孕母鼠的输卵管内，９只母鼠妊娠，共生３７只小鼠。用Ｓｏｕｔｈｅｒｎ和Ｎｏｒｔｈｅｒｎ杂交法检测外源基因在小鼠染色体的整合与晶体的表达情况，Ｅ＿２号鼠为有整合、有ｍＲＮＡ表达的转基因小鼠。在人老年性白内障晶体，Ｓｏｕｔｈｅｒｎ杂交显示ＢａｍＨＩ－ｖｉｍｅｎｔｉｎ基因片段较透明晶体对照组信号明显增强。

To investigate the role of vimentin gene in cataractogenesis,12.7 Kb chicken vimentin genes were microinjected into the male pronuclei of 530 fertilized mouse eggs. 453 injected embryos were transferred into the oviducts of 18 pseudopregnant recipient fernales,of which 9 pregnant mice gave blrths to 37 offspring mice.The integration and expression of exogenous gene in the offsprings were an-alyzed by Southern and Northern blot hybridizations.It is shown that the E_2 offspring was a transgenic mouse in which the chicken vimentin gene was integrated in its genome and efficiently expressed in mRNA level in its lens.In the human senile cataract lens,Bam-HI-restricted vimentin fragments dis-played a stronger hybridization signal than that of the control lens in Southern blot analysis.