Wei Y G, Surzycki S J
Gene. 1986;48(2-3):251-6. doi: 10.1016/0378-1119(86)90083-1.
Detection and isolation of Escherichia coli clones carrying vectors with foreign DNA sequences partially homologous to specific E. coli genes is difficult because denatured DNA in the host genome can hybridize with the probe. In this paper we present a procedure which simplifies this task by using bacteriophage M13 as the cloning vector. The procedure takes advantage of the secretory properties of the phage, as well as the property of nitrocellulose membrane to bind protein and single-stranded DNA but not double-stranded DNA. This procedure is shown to be effective in identifying E. coli clones containing sequences of Chlamydomonas reinhardtii chloroplast DNA that are homologous to the rpoC gene of E. coli. We suggest that this procedure can be used generally for rapid isolation of DNA sequences that are homologous to E. coli genes.
携带与特定大肠杆菌基因部分同源的外源DNA序列载体的大肠杆菌克隆的检测与分离很困难，因为宿主基因组中的变性DNA会与探针杂交。在本文中，我们提出了一种通过使用噬菌体M13作为克隆载体来简化这项任务的方法。该方法利用了噬菌体的分泌特性，以及硝酸纤维素膜结合蛋白质和单链DNA而不结合双链DNA的特性。结果表明，该方法对于鉴定含有与大肠杆菌rpoC基因同源的莱茵衣藻叶绿体DNA序列的大肠杆菌克隆是有效的。我们认为，该方法一般可用于快速分离与大肠杆菌基因同源的DNA序列。
