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用特定DNA片段进行的大核转化控制了四膜虫新大核基因组的含量。

Macronuclear transformation with specific DNA fragments controls the content of the new macronuclear genome in Paramecium tetraurelia.

作者信息

You Y, Aufderheide K, Morand J, Rodkey K, Forney J

机构信息

Department of Biochemistry, Purdue University, West Lafayette, Indiana 47907.

出版信息

Mol Cell Biol. 1991 Feb;11(2):1133-7. doi: 10.1128/mcb.11.2.1133-1137.1991.

Abstract

A previously isolated mutant cell line called d48 contains a complete copy of the A surface antigen gene in the micronuclear genome, but the gene is not incorporated into the macronucleus. Previous experiments have shown that a cytoplasmic factor made in the wild-type macronucleus can rescue the mutant. Recently, S. Koizumi and S. Kobayashi (Mol. Cell. Biol. 9:4398-4401, 1989) observed that injection of a plasmid containing the A gene into the d48 macronucleus rescued the cell line after autogamy. It is shown here that an 8.8-kb EcoRI fragment containing only a portion of the A gene coding region is sufficient for the rescue of d48. The inability of other A gene fragments to rescue the mutant shows that this effect is dependent upon specific Paramecium DNA sequences. Rescue results in restoration of the wild-type DNA restriction pattern in the macronucleus. These results are consistent with a model in which the macronuclear A locus normally makes an additional gene product that is required for correct processing of the micronuclear copy of the A gene.

摘要

一种先前分离出的名为d48的突变细胞系,其微核基因组中含有完整的A表面抗原基因拷贝,但该基因未整合到大核中。先前的实验表明,野生型大核中产生的一种细胞质因子可以挽救该突变体。最近,小泉S.和小林S.(《分子细胞生物学》9:4398 - 4401,1989年)观察到,将含有A基因的质粒注入d48大核后,在自体受精后挽救了该细胞系。本文表明,一个仅包含A基因编码区一部分的8.8 kb EcoRI片段足以挽救d48。其他A基因片段无法挽救该突变体,这表明这种效应取决于特定的草履虫DNA序列。挽救导致大核中野生型DNA限制图谱的恢复。这些结果与一个模型一致,在该模型中,大核A位点通常产生一种额外的基因产物,该产物是正确处理A基因微核拷贝所必需的。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8eb3/359794/29b37c7f22f3/molcellb00137-0561-a.jpg

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