Herzog R W, Singh N K, Schmidt C, Lemke P A
Department of Botany and Microbiology, Auburn University, AL 36849, USA.
Curr Genet. 1995 Apr;27(5):460-5. doi: 10.1007/BF00311216.
Replicative plasmids pP01 and pP02, recovered from Pleurotus ostreatus transformants, contain an insert of bacteriophage origin. These plasmids have been amplified by the polymerase chain reaction (PCR) and have been shown to represent a low-grade component in the initial preparation of the vector pAN7-1. The pP01 and pP02 plasmids share an insert (P01A) of virtual identity with a SmaI-BamHI genomic fragment of P 1 bacteriophage and retain remnants of a polylinker at the 5' end of this fragment. Such an insert undoubtedly represents an in vitro-generated event and did not arise, as suggested previously, by recombination of pAN7-1 with the P. ostreatus genome. The P. ostreatus transformants, however, do select for the minority pP0 plasmid, apparently recognizing the P01A insert as a heterologous or surrogate replicon.
从糙皮侧耳转化体中回收的复制性质粒pP01和pP02含有噬菌体起源的插入片段。这些质粒已通过聚合酶链反应(PCR)进行扩增,并已证明在载体pAN7-1的初始制备中代表一种低水平成分。pP01和pP02质粒与P1噬菌体的SmaI-BamHI基因组片段共享一个几乎完全相同的插入片段(P01A),并在该片段的5'端保留了多克隆位点的残余部分。这样的插入片段无疑代表了一个体外产生的事件,并非如先前所暗示的那样,由pAN7-1与糙皮侧耳基因组重组产生。然而,糙皮侧耳转化体确实选择了少数的pP0质粒,显然将P01A插入片段识别为异源或替代复制子。