Chen X J, Wésolowski-Louvel M, Tanguy-Rougeau C, Fukuhara H
Institut Curie, Section de Biologie, Orsay, France.
Biochimie. 1991 Sep;73(9):1195-203. doi: 10.1016/0300-9084(91)90004-k.
The killer plasmid k1 of Kluyveromyces lactis has terminal inverted repeats of 202 base pairs (bp). The left terminal repeat is contiguous to the transcribed open reading frame, ORF1, which is supposed to code for a DNA polymerase. A 266-bp fragment (called Pk1) containing most of the terminal repeat sequence was isolated and examined for promoter activity. Pk1 was fused, in either original or inversed orientation, with a promoter-less lacZ gene of E coli and a promoter-less G418 resistance gene of Tn903. These fusions were introduced into a pKD1-derived circular vector, and transformed into a lactose-negative (lac4), and a G418-sensitive K lactis host. Lac+ and G418-resistant transformants were obtained with either orientation of Pk1. The promoter activity of Pk1 fragment was independent of the presence or absence of killer plasmids. It is not known whether Pk1 can also function bidirectionally on the natural k1 plasmid. The possible functions of Pk1 for killer plasmid gene expression and plasmid replication are discussed.
乳酸克鲁维酵母的杀伤性质粒k1具有202个碱基对(bp)的末端反向重复序列。左侧末端重复序列与转录的开放阅读框ORF1相邻,该开放阅读框被认为编码一种DNA聚合酶。分离出一个包含大部分末端重复序列的266 bp片段(称为Pk1),并检测其启动子活性。Pk1以原始或反向方向与大肠杆菌无启动子的lacZ基因和Tn903的无启动子G418抗性基因融合。将这些融合体导入源自pKD1的环状载体中,并转化到乳糖阴性(lac4)和对G418敏感的乳酸克鲁维酵母宿主中。无论Pk1的方向如何,都获得了Lac+和G418抗性转化体。Pk1片段的启动子活性与杀伤性质粒的存在与否无关。尚不清楚Pk1在天然k1质粒上是否也能双向发挥作用。本文讨论了Pk1在杀伤性质粒基因表达和质粒复制中的可能功能。