Fierro F, Kosalková K, Gutiérrez S, Martin J F
Area of Microbiology, Faculty of Biology, University of León, and Institute of Biotechnology, INBIOTEC, E-24071 León, Spain.
Curr Genet. 1996 Apr;29(5):482-9. doi: 10.1007/BF02221518.
Plasmid vectors containing the AMA1 sequence transformed with high efficiency and replicated autonomously in Penicillium chrysogenum. The efficiency of transformation of P. chrysogenum was related to the length of the AMA1 fragment used for constructing the different autonomously replicating plasmids. One of the two palindromic inverted repeats of AMA1 (the 2.2-kb SalI-HindIII fragment) is sufficient to confer autonomous replication and a high transformation efficiency. Deletion of the 0.6-kb central fragment located between the inverted repeats did not affect either the ability of the plasmids to replicate autonomously or the efficiency of transformation, but did alter the mitotic stability and the plasmid copy number. Deletion of any fragment of the 2.2-kb repeat caused the loss of the ability to replicate autonomously and reduced the transformation efficiency. Most of the transformants retained the original plasmid configuration, as multimers and without reorganization, after several rounds of autonomous replication. The AMA1 region works as an origin of replication in P. chrysogenum and A. nidulans but not apparently in Acremonium chrysogenum.
含有AMA1序列的质粒载体在产黄青霉中能高效转化并自主复制。产黄青霉的转化效率与用于构建不同自主复制质粒的AMA1片段长度有关。AMA1的两个回文反向重复序列之一(2.2 kb的SalI - HindIII片段)足以赋予自主复制能力和高转化效率。位于反向重复序列之间的0.6 kb中央片段的缺失既不影响质粒自主复制的能力,也不影响转化效率,但会改变有丝分裂稳定性和质粒拷贝数。2.2 kb重复序列的任何片段的缺失都会导致自主复制能力的丧失并降低转化效率。经过几轮自主复制后,大多数转化体保留了原始质粒构型,呈多聚体且无重组。AMA1区域在产黄青霉和构巢曲霉中作为复制起点起作用,但在产黄顶孢霉中显然不起作用。