Ebe K, Schöld M, Rossi J J, Wallace R B
Department of Molecular Biochemistry, Beckman Research Institute of the City of Hope, Duarte, CA 91010.
DNA. 1987 Oct;6(5):497-504. doi: 10.1089/dna.1987.6.497.
A plasmid DNA vector is described that is suitable for cloning synthetic DNA sequences. These cloned synthetic DNA sequences can be transcribed in vitro to produce oligoribonucleotides of defined sequence. Transcription is directed by a promoter based on the consensus sequence for Escherichia coli promoters and uses E. coli RNA polymerase. The vector is useful for cloning oligodeoxyribonucleotides of mixed sequences, the individual sequences being resolved by transformation and colony selection. Oligoribonucleotide synthesis from the vector is highly specific. Application of these sequences in hybridization experiments is demonstrated.
描述了一种适用于克隆合成DNA序列的质粒DNA载体。这些克隆的合成DNA序列可在体外转录以产生确定序列的寡核糖核苷酸。转录由基于大肠杆菌启动子共有序列的启动子指导,并使用大肠杆菌RNA聚合酶。该载体可用于克隆混合序列的寡脱氧核糖核苷酸,通过转化和菌落选择来解析各个序列。从该载体合成寡核糖核苷酸具有高度特异性。展示了这些序列在杂交实验中的应用。