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秀丽隐杆线虫的染色体外DNA转化

Extrachromosomal DNA transformation of Caenorhabditis elegans.

作者信息

Stinchcomb D T, Shaw J E, Carr S H, Hirsh D

出版信息

Mol Cell Biol. 1985 Dec;5(12):3484-96. doi: 10.1128/mcb.5.12.3484-3496.1985.

Abstract

DNA was introduced into the germ line of the nematode Caenorhabditis elegans by microinjection. Approximately 10% of the injected worms gave rise to transformed progeny. Upon injection, supercoiled molecules formed a high-molecular-weight array predominantly composed of tandem repeats of the injected sequence. Injected linear molecules formed both tandem and inverted repeats as if they had ligated to each other. No worm DNA sequences were required in the injected plasmid for the formation of these high-molecular-weight arrays. Surprisingly, these high-molecular-weight arrays were extrachromosomal and heritable. On average 50% of the progeny of a transformed hermaphrodite still carried the exogenous sequences. In situ hybridization experiments demonstrated that approximately half of the transformed animals carried foreign DNA in all of their cells; the remainder were mosaic animals in which some cells contained the exogenous sequences while others carried no detectable foreign DNA. The presence of mosaic and nonmosaic nematodes in transformed populations may permit detailed analysis of the expression and function of C. elegans genes.

摘要

通过显微注射将DNA导入线虫秀丽隐杆线虫的生殖系。大约10%的注射线虫产生了转化后代。注射后,超螺旋分子形成了一种高分子量阵列,主要由注射序列的串联重复组成。注射的线性分子形成了串联和反向重复,就好像它们彼此连接在一起一样。在注射的质粒中形成这些高分子量阵列不需要线虫DNA序列。令人惊讶的是,这些高分子量阵列是染色体外的且可遗传。平均而言,转化雌雄同体的后代中有50%仍然携带外源序列。原位杂交实验表明,大约一半的转化动物在其所有细胞中都携带外源DNA;其余的是嵌合体动物,其中一些细胞含有外源序列,而另一些细胞则没有可检测到的外源DNA。转化群体中嵌合体和非嵌合体线虫的存在可能有助于对线虫基因的表达和功能进行详细分析。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d708/369179/67467a917332/molcellb00142-0167-a.jpg

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