INTEGRATIVE TRANSFORMATION OF CAENORHABDITIS-ELEGANS
INTEGRATIVE TRANSFORMATION OF CAENORHABDITIS-ELEGANS
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DOI：
10.1002/j.1460-2075.1986.tb04550.x
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发表时间：
1986-10-01
期刊：
影响因子：
11.4
通讯作者：
FIRE, A
中科院分区：
文献类型：
--
作者：
FIRE, A
介绍了一种将外源DNA导入秀丽线虫染色体的技术。利用克隆的线虫琥珀抑制因子tRNA基因sup-7作为选择标记。这种琥珀抑制子的活性是通过注射携带琥珀终止突变基因(tra-3)的蠕虫来选择的，该基因的功能是生育所必需的。在注射后的一代中，可育(被拯救)的动物的存在证明了sup-7的瞬时表达。在少数情况下，这些可育动物产生了稳定的抑制系(到目前为止已经确定了8个)。每一条稳定的抑制子都携带注入的DNA序列。抑制活性已被定位在染色体上，表明外源DNA已整合到基因组中。这项技术已被用于引入含有果蝇热休克启动子元件的嵌合基因，该嵌合基因与大肠杆菌β-半乳糖苷酶基因的编码序列融合在一起。这种嵌合基因在稳定转化的品系中起作用，并且是热诱导的。
A technique for introducing exogenous DNA into the chromosomes of the nematode Caenorhabditis elegans is presented. A cloned C. elegans amber suppressor tRNA gene, sup-7, is used as a selective marker. The activity of this amber suppressor is selected for by injecting worms which carry an amber termination mutation in a gene (tra-3) whose function is required for fertility. Transient expression of sup-7 is evidenced by the presence of fertile (rescued) animals in the generation after injection. In a fraction of cases, these fertile animals give rise to stable suppressor lines (eight have been characterized so far). Each of the stable suppressor lines carries injected DNA sequences. The suppressor activities have been mapped to chromosomal loci, indicating that the exogenous DNA has integrated into the genome. This technique has been used to introduce a chimeric gene containing a Drosophila heat shock promoter element fused to coding sequences from the Escherichia coli .beta.-galactosidase gene. This chimeric gene functions and is heat inducible in the resulting stably transformed lines.