An improved vector for the expression of proteins in all three translational reading frames.
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.
Seth
A
A
Laboratory of Molecular Oncology, National Cancer Institute, Frederick, MD 21701-1013.
Thompson
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D
Chen
S M
SM
Papas
T S
TS
D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods; D003062:Codon / Q000096:biosynthesis; D004247:DNA / Q000302:isolation & purification; D011088:DNA Ligases; D010957:Plasmids; D011401:Promoter Regions, Genetic; D014176:Protein Biosynthesis; D014157:Transcription Factors / Q000096:biosynthesis / Q000235:genetics*; D014764:Viral Proteins
D003062:Codon; D014157:Transcription Factors; D014764:Viral Proteins; C039000:cII protein, bacteriophage lambda; D004247:DNA; D011088:DNA Ligases