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曹月青 周玲 韩柱 尚磊雄 虎良菊 姚文彬CAO Yue-qing ZHOU Ling HAN Zhu SHANG Lei-xiong HU Liang-ju YAO Wen-bin 《遗传》2002,24(4):407-165
利用定点突变人胰岛素基因,以脂质体载体和质粒不同比例形成的复合物进行体内外转染。体外转染鼠肝细胞,G418进行筛选,利用放免法测定培养液中胰岛素含量;体内通过肝门脉注射,测定模型鼠血糖及转染后7天时血液中胰岛素的含量,结果显示目的基因已转入肝细胞,且体内外转染都有一定量的成熟胰岛素表达,体外转染中质粒与脂质体比为1∶6转染后24h表达量最高为10.45μIU/ml,体内转染使模型鼠的糖尿病症状明显改善,血糖最高降幅达55%。
Abstract:The transfection of mutated human insulin gene was studied using the complex of different proportion of plasmid and Liposome.Hepatic cell was used as the target cell in vitro,isolation of Hepatic cell including insulin gene was carried out by G418,the expression level of insulin in medium was measured by RIA method.The portal vein was cannulated with therapeutic gene in vivo,the blood glucose of the model was regularly examined and the insulin level was detected on the seventh day after transfection.The results showed that the target gene was transferred into the hepatic cell,expression of mature insulin was detected both in vivo and in vitro,It reached the peak 10.45μIU/ml on the 24th hour after transfection with the proportion 1:6 of.plasmid and Liposome in hepatic cell.Diabetic sympotom of the model was improved after transgene,the blood glucose could decrease 55% at the most. 相似文献
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植物β-胡萝卜素羟化酶(简称β-羟化酶)是玉米黄素合成过程中的关键酶。柑橘、拟南芥、大白菜等多种植物β-羟化酶基因都已被克隆。柑橘中合成玉米黄素的中间产物β-隐黄素的含量远远高于其它植物,推测柑橘与其它植物的β-羟化酶基因相比有一些特殊的差异。本研究以南丰蜜橘为材料,克隆了柑橘β-羟化酶全长基因,并对该基因内含子进行定位和分析,结果表明南丰蜜橘β-羟化酶基因具有6个内含子,其中内含子6的长度远大于拟南芥和大白菜中相应的内含子。进一步研究发现内含子6中有两段特殊序列,一段长122 bp,另一段长95 bp。特别是122 bp序列两端具有反向重复序列,全序列中含有许多不同的转录起始原件。二级结构预测其对应RNA二级结构能构形成较稳定的发夹结构。 相似文献
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