首页> 外文期刊>The Indian Journal of Animal Reproduction >EVALUATION OF TRANSFECTED FIBROBLAST CELLS OF SAHIWAL CATTLE FOR TRANSFECTION / NUCLEAR TRANSFER STUDIES
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EVALUATION OF TRANSFECTED FIBROBLAST CELLS OF SAHIWAL CATTLE FOR TRANSFECTION / NUCLEAR TRANSFER STUDIES

机译:沙伊瓦尔牛转化成纤维细胞进行核/核转移的评价。

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摘要

The development of an efficient transfection system in livestock cells is an important step towards investigating gene transfer and has impact mainly in agriculture and fundamental biology. The developmental capacity of transfected fibroblast cells isolated from skin piece of ear pinna of adult Sahiwal cattle was studied. Effect of transfection and in vitro culture of cells for six successive passages post transfection was evaluated. Important factors involved in cationic liposome mediated gene transfer were evaluated through in vitro transfection of bovine fibroblast cells. Transfection of plasmid DNA complex (pEGFP and DNAfectin reagent) was evaluated and standardized utilizing the parameters concentration of DNA, lipofection reagent, cell density, and the effect of transfection time on the efficiency of indigenous Sahiwal cattle fibroblasts to express a reporter gene. Stable transfections were conducted using a green fluorescence protein (GFP) reporter plasmid containing the neomycin resistancegene.
机译:牲畜细胞中高效转染系统的开发是研究基因转移的重要一步,主要影响农业和基础生物学。研究了从成年Sahiwal牛耳廓皮中分离的转染的成纤维细胞的发育能力。评估转染后六次连续转染和细胞体外培养的效果。通过体外转染牛成纤维细胞评估了阳离子脂质体介导的基因转移的重要因素。评估质粒DNA复合物(pEGFP和DNAfectin试剂)的转染,并利用DNA的浓度,脂转染试剂,细胞密度以及转染时间对Sahiwal牛成纤维细胞表达报告基因效率的影响进行标准化。使用包含新霉素抗性基因的绿色荧光蛋白(GFP)报告质粒进行稳定的转染。

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