Gene transfer for targeted modification of salmonid fish metabolism

Aleksei Krasnov, Tiina I Pitkänen, Hannu Mölsä
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引用次数: 14

Abstract

The reviewed studies addressed the possibility of using gene transfer for correction of l-ascorbic acid biosynthesis and carbohydrate utilization in rainbow trout. Analyses of enzymatic activities in the l-AAB pathway indicated that reasons for the lack of l-AA production can be common in fish and scurvy-prone animals. Rat gulonolactone oxidase cDNA was transferred into trout. Regardless of the fact that rGLO transcription occurred in embryos, neither GLO protein, nor enzyme activity were detected. There was no production of l-AA in transgenic fish raised on vitamin C-free diets or injected with l-gulonolactone. These results indicated that the conditions required for translation or stability of rGLO were not present in trout tissues. To augment carbohydrates utilization, human glucose transporter 1 and rat hexokinase II cDNAs were tested. In the transfected embryos, HK activity, rates of hexose uptake and glucose oxidation were increased. The effect of hGLUT1 on glucose metabolism was greater than that of rHKII. Trout carrying hGLUT1 and rHKII with viral or piscine promoters were created. Though interpretation of the metabolic effects of the transgenes was complicated with mosaicism, a tendency to improved carbohydrate utilization was revealed in some of the transgenic individuals.

靶向修饰鲑鱼代谢的基因转移
综述了利用基因转移校正虹鳟鱼l-抗坏血酸生物合成和碳水化合物利用的可能性。l-AAB途径的酶活性分析表明,在鱼类和容易患坏血病的动物中,缺乏l-AA的原因可能是共同的。将大鼠古罗内酯氧化酶cDNA转移到鳟鱼体内。尽管rGLO转录在胚胎中发生,但GLO蛋白和酶活性均未检测到。无维生素c饲料饲养和注射l-古洛内酯的转基因鱼没有产生l-AA。这些结果表明,rGLO在鳟鱼组织中不存在翻译或稳定所需的条件。为了增加碳水化合物的利用,我们检测了人葡萄糖转运蛋白1和大鼠己糖激酶II cdna。在转染后的胚胎中,HK活性、己糖摄取率和葡萄糖氧化率均有所增加。hGLUT1对糖代谢的影响大于rHKII。鳟鱼携带hGLUT1和rHKII与病毒或鱼类启动子。虽然对转基因的代谢效应的解释与嵌合现象是复杂的,但在一些转基因个体中显示出改善碳水化合物利用的趋势。
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