Effect of de-phosphorylation of linearized pAN7-1 and of addition of restriction enzyme on plasmid integration in Penicillium paxilli

被引:17
作者
Itoh, Y [1 ]
Scott, B [1 ]
机构
[1] MASSEY UNIV, DEPT GENET & MICROBIOL, MOL GENET UNIT, PALMERSTON NORTH, NEW ZEALAND
基金
日本学术振兴会;
关键词
Penicillium paxilli; plasmid integration; homologous recombination; restriction enzyme-mediated (REM) integration;
D O I
10.1007/s002940050259
中图分类号
Q3 [遗传学];
学科分类号
071007 ; 090102 ;
摘要
As part of a study to investigate the pathways of plasmid pAN7-1 integration in Penicillium paxilli, a molecular analysis of 90 different integration events was carried out. Twenty out of forty five integration events analyzed from transformants obtained without the addition of restriction enzyme to the transformation reaction mixture were single-copy integrations, whereas the remaining 25 were tandem-repeat integrations, The addition of restriction enzyme resulted in a shift in this ratio in favour of single-copy integration events. Analysis of the 33 tandem-repeat integration events showed that the orientation of the plasmid copies was not random, with 88% organized as tandem head-to-tail arrays. De-phosphorylation of linearized pAN7-1 did not affect the frequency with which multiple copies were integrated. This suggests that the predominant mechanism for the generation of tandem repeats in P. paxilli is by homologous recombination rather than in vivo ligation of linearized plasmids.
引用
收藏
页码:147 / 151
页数:5
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