Seth A, Thompson D, Chen S M, Papas T S
Laboratory of Molecular Oncology, National Cancer Institute, Frederick, MD 21701-1013.
Gene Anal Tech. 1989 Sep-Oct;6(5):97-100. doi: 10.1016/0735-0651(89)90015-0.
We have constructed a novel vector (pN-7) that is capable of producing large amounts of recombinant proteins in E. coli and requires minimal manipulation for the construction of recombinant expression vectors. This expression vector (pN-7) contains the tightly regulated lambda pL promoter, cII ribosome binding site, and initiator condon ATG. The pN-7 vector also contains cleavage sites for the restriction enzymes SmaI, EcoRV, and HpaI that provide blunt ends in all three reading frames. Thus after cleavage with the appropriate restriction enzyme, this novel vector can be directly ligated to the DNA fragment that contains the open reading frame without further manipulation.
我们构建了一种新型载体(pN - 7),它能够在大肠杆菌中大量产生重组蛋白,并且构建重组表达载体所需的操作极少。这种表达载体(pN - 7)包含严格调控的λ pL启动子、cII核糖体结合位点和起始密码子ATG。pN - 7载体还包含限制性内切酶SmaI、EcoRV和HpaI的切割位点,这些位点在所有三个阅读框中都提供平端。因此,用适当的限制性内切酶切割后,这种新型载体可以直接与包含开放阅读框的DNA片段连接,无需进一步操作。