TY - JOUR
T1 - A plasmid expression vector that permits stabilization of both mRNAs and proteins encoded by the cloned genes
AU - Duvoisin, Robert M.
AU - Belin, Dominique
AU - Krisch, Henry M.
N1 - Funding Information:
We are grateful to S. Frutiger, S. Gasser, U. Laemmli,T . Mattson and P. Prentko for helpful discussionsa, ndt o R. Epstein,J .-C. Jaton andJ .-D. Vassalli for aid, encouragemenatn d support.W e thankB . Allet (BiogenS .A., Geneva)f or the gift of plasmidD NA, 0. Jenni for preparingt he illustra-tionsa ndR. Bisig andM . Burgatf or skillfula idwith someo f the experimentsT.h is work was supported in part by grants (3.411-0.83,3 .075-0.84a nd 3.465.83) from the Swiss National Science Foundation.A dditional supporta nd facilitiesw ere providedb y theD epartmenot f Public Instructiono f the Stateo f Geneva.
PY - 1986
Y1 - 1986
N2 - Two new expression vectors have been constructed to take advantage of several useful properties of bacteriophage T4-infected Escherichia coli.These plasmids,pRDB8 and pRDB9,contain the promoter region and start codon of T4 gene 32, a contiguous multiple cloning site (MCS),and translation and transcription termination signals. DNA fragments inserted into the MCS are transcribed and translated at a high level in both uninfected and phage T4-infected cells.Furthermore, the extreme stability of the hybrid mRNA after infection permits the specific biosynthetic labeling of the protein encoded by the cloned gene.In addition, the cloned gene product is stabilized, since the host-mediated degradation of foreign proteins is inhibited by phage infection. The properties of this expression system were demonstrated with the constant region of a rabbit immunoglobulin λ light chain (Cλ) gene.Although proteolytic degradation of the Cλfusion protein was rapid in uninfected cells,degradation was blocked in phage-infected cells and the protein accumulated in greater amounts.
AB - Two new expression vectors have been constructed to take advantage of several useful properties of bacteriophage T4-infected Escherichia coli.These plasmids,pRDB8 and pRDB9,contain the promoter region and start codon of T4 gene 32, a contiguous multiple cloning site (MCS),and translation and transcription termination signals. DNA fragments inserted into the MCS are transcribed and translated at a high level in both uninfected and phage T4-infected cells.Furthermore, the extreme stability of the hybrid mRNA after infection permits the specific biosynthetic labeling of the protein encoded by the cloned gene.In addition, the cloned gene product is stabilized, since the host-mediated degradation of foreign proteins is inhibited by phage infection. The properties of this expression system were demonstrated with the constant region of a rabbit immunoglobulin λ light chain (Cλ) gene.Although proteolytic degradation of the Cλfusion protein was rapid in uninfected cells,degradation was blocked in phage-infected cells and the protein accumulated in greater amounts.
KW - Recombinant DNA
KW - bacteriophage T4 gene 32
KW - immunoglobulin lambda chain
KW - protease inhibition
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U2 - 10.1016/0378-1119(86)90254-4
DO - 10.1016/0378-1119(86)90254-4
M3 - Article
C2 - 3026907
AN - SCOPUS:0023017879
SN - 0378-1119
VL - 45
SP - 193
EP - 201
JO - Gene
JF - Gene
IS - 2
ER -