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文档为南京师范大学生命科学学院 09 级《基 因工程》课程作业,翻译一篇英文论文。此为本 人亲自翻译。仅供参考!
Retrovirus-mediated gene transfer and expression cloning: Powerful tools in functional genomics
Most of the human genome has now been sequenced and about 30,000 potential open reading frames have been identified, indicating that we use these 30,000 genes to functionally organize our biologic activities. However, functions of many genes are still unknown despite intensive efforts using bioinformatics as well as transgenic and knockout mice. Retrovirus-mediated gene transfer is a powerful tool that can be used to understand gene functions. We have developed a variety of retrovirus vectors and efficient packaging cell lines that have facilitated the development of efficient functional expression cloning methods. In this review, we describe retrovirus-mediated strategies used for investigation of gene functions and function-based screening strategies. 2003 International Society for Experimental Hematology.Published by Elsevier Inc.
摘要:人类基因组的大部分现在已经测序完成,大约 30,000 潜在的开放阅读框已经确 定,表明我们使用这 30,000 个基因管理我们的生物学活和功能性。然而,许多基因的 功能仍是未知尽管在生物信息学上以及转基因和基因敲除小鼠做出了大量的努力工作。 逆转录病毒介导的基因转移是一个功能强大的工具,可以用来了解基因的功能。我们已 经开发出多种逆转录病毒载体和高效的包装细胞系, 有利于发展高效功能表达克隆的方 法。本文中,我们描述了逆转录病毒介导的方法,用于基因功能和功能为基础的筛选策 略的研究。 2003 International Society for Experimental Hematology.Published by
Elsevier Inc.
Function-based gene cloning It was only 30 years ago that recombinant DNA technology was initiated [1]. The globin gene was cloned as the first mammalian gene in 1976 that was reverse transcribed from the purifiedmRNAfor globin [2]. For less abundant mRNAs, the cDNAs were frequently isolated based on the amino acid sequences of purified proteins. In late 1970s, a method called hybrid selection was developed. The principle of this method was to detect proteins translated from the mRNA hybridized to a particular pool of subdivided cDNA library fixed on nitrocellulose membranes, thereby identifying a pool that contains a cDNA of interest. Xenopus o
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