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[从莱茵衣藻杂种自主质粒中分离的染色体DNA片段的分析]

[Analysis of a chromosomal DNA fragment isolated from a hybrid autonomous plasmid of Chlamydomonas reinhardtii].

作者信息

Sizova I A, Lapina T V, Kozlov A V

出版信息

Genetika. 1995 Mar;31(3):324-32.

PMID:7607422
Abstract

The ability of a previously cloned fragment of nuclear DNA of Chlamydomonas reinhardtii (a 2.4-kb BamH 1-EcoR I fragment) to function as the origin of replication of plasmid pARG7.8.2.4, carrying the homologous arg7 marker gene of prototrophy or arginine in cells of C. reinhardtii transformants, was studied. As shown by Southern blotting, plasmid pARG7.8.2.4 integrates at several sites of chromosomal DNA of chlamydomonas without autonomous maintenance in cells. Results obtained using Southern blotting and plasmid rescue techniques demonstrated that plasmid pARG7.8, containing only the arg7 gene, is able both to integrate into the chromosome and to be maintained, in some cases, in the autonomous state in Arg+ transformants of alga. Analysis of the nucleotide sequence of the DNA fragment under study revealed several types of GC-enriched repeats that manifested homology with the nucleotide sequence of the arg7 gene in introns 10 and 12.

摘要

研究了莱茵衣藻核DNA的一个先前克隆片段(一个2.4 kb的BamH 1-EcoR I片段)作为质粒pARG7.8.2.4复制起点的功能,该质粒携带莱茵衣藻转化体细胞中原养型或精氨酸的同源arg7标记基因。Southern印迹分析表明,质粒pARG7.8.2.4整合到衣藻染色体DNA的多个位点,在细胞中不能自主维持。使用Southern印迹和质粒拯救技术获得的结果表明,仅含有arg7基因的质粒pARG7.8既能整合到染色体中,在某些情况下,又能在藻类的Arg+转化体中以自主状态维持。对所研究DNA片段的核苷酸序列分析揭示了几种富含GC的重复序列类型,这些序列与内含子10和12中arg7基因的核苷酸序列表现出同源性。

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