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黑腹果蝇琥珀抑制tRNA(Ser)在秀丽隐杆线虫中的表达。

Expression of a Drosophila melanogaster amber suppressor tRNA(Ser) in Caenorhabditis elegans.

作者信息

Pilgrim D B, Bell J B

机构信息

Department of Genetics, University of Alberta, Edmonton, Canada.

出版信息

Mol Gen Genet. 1993 Oct;241(1-2):26-32. doi: 10.1007/BF00280197.

Abstract

The purpose of this study was to test a cloned amber-suppressing tRNA(Ser) gene derived from Drosophila melanogaster for its ability to produce amber suppression in the nematode Caenorhabditis elegans. To date, all characterized nonsense suppressors in C. elegans have been derived from tRNA(Trp) genes. Suppression was assayed by monitoring the reversal of a mutant tra-3 phenotype among individuals transformed with the cloned Drosophila suppressor gene. An amber allele of tra-3 results in masculinization of XX animals with accompanying sterility. Complete suppression was observed among the transformants. The presence of the heterologous transgene, in both suppressed experimental animals and controls injected with a non-suppressing wild-type Drosophila tRNA(Ser) gene, was verified by PCR amplification of DNA from single worms using primers flanking the tRNA(Ser) gene. Suppression by the heterologous transgene was comparable in quality to that produced by endogenous C. elegans suppressors, and, in frequency as well as quality, to that produced by a transgenic C. elegans tRNA(Trp)-derived suppressors. Thus, a heterologous suppressor gene will function in C. elegans, and it need not be based on tRNA(Trp).

摘要

本研究的目的是测试一个源自黑腹果蝇的克隆琥珀抑制tRNA(Ser)基因在秀丽隐杆线虫中产生琥珀抑制的能力。迄今为止,秀丽隐杆线虫中所有已鉴定的无义抑制子均源自tRNA(Trp)基因。通过监测用克隆的果蝇抑制基因转化的个体中突变tra-3表型的逆转来检测抑制作用。tra-3的一个琥珀等位基因导致XX动物雄性化并伴有不育。在转化体中观察到完全抑制。通过使用位于tRNA(Ser)基因两侧的引物对单条线虫的DNA进行PCR扩增,验证了在受抑制的实验动物和注射了非抑制性野生型果蝇tRNA(Ser)基因的对照中异源转基因的存在。异源转基因产生的抑制在质量上与秀丽隐杆线虫内源性抑制子产生的抑制相当,在频率和质量上也与转基因秀丽隐杆线虫tRNA(Trp)衍生的抑制子产生的抑制相当。因此,一个异源抑制基因将在秀丽隐杆线虫中发挥作用,并且它不必基于tRNA(Trp)。

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