Tacon W C, Bonass W A, Jenkins B, Emtage J S
Gene. 1983 Sep;23(3):255-65. doi: 10.1016/0378-1119(83)90016-1.
Two DNA fragments which contain the Escherichia coli tryptophan promoter operator region but lack the attenuator have been used in the construction of a series of pAT153 based plasmids suitable for the regulated expression of foreign genes in E. coli. The first, a 139-bp HhaI fragment includes 59 bp of the trp leader sequence, ending within the "attenuator peptide" coding sequence, eleven codons from the N-terminus. A fusion-type expression plasmid incorporating this fragment has been constructed. The second, a 99-bp HaeIII-TaqI fragment contains no coding sequence but includes the "attenuator peptide" SD site situated 4 bp upstream of the TaqI site. This fragment has been incorporated in expression vectors which result in the direct expression of cloned gene sequences. To further maximise expression, plasmids with directly repeating trp promoter HaeIII-TaqI units have been constructed.
两个含有大肠杆菌色氨酸启动子操纵区但缺少弱化子的DNA片段已被用于构建一系列基于pAT153的质粒,这些质粒适合在大肠杆菌中对外源基因进行调控表达。第一个是139 bp的HhaI片段,包括59 bp的trp前导序列,终止于“衰减肽”编码序列内,距N端11个密码子处。已构建了一个包含该片段的融合型表达质粒。第二个是99 bp的HaeIII-TaqI片段,不含编码序列,但包括位于TaqI位点上游4 bp处的“衰减肽”SD位点。该片段已被整合到表达载体中,可直接表达克隆的基因序列。为了进一步最大化表达,已构建了具有直接重复trp启动子HaeIII-TaqI单元的质粒。