Biopolym. Cell. 1986; 2(5):275-278.
Short Communications
Insertion of a DNA fragment between two strong similarly oriented promoters of recombinant filamentous phage mp8t0/rpoB increases its stability and makes possible an opposite orientation of the cloned E. coli rpoB gene
- Institute of Molecular Biology and Genetics, Academy of Sciences of the Ukrainian SSR
Kiev, USSR
Abstract
A fragment of λ phage DNA containing terminator of transcription (t0) and terminal part of the oop-RNA gene was inserted into the polylinker area of M13 mp8 filamentous phage. The obtained phage was used to clone a BglII fragment of pJC703 cosmid, containing E. coli genes rpIJ, rpIL and rpoB together with promoters PJ and Pβ. Stability of the obtained mp8t /rpoB recombinant phages increased up to 93 % and an orientation of the cloned BglII-fragment opposite to the previously observed one became possible.
Full text: (PDF, in Russian)
References
[1]
Paton EB, Vudmaska MI, Sverdlov ED. Unidirectional orientation of the rpo B gene of E. coli cloned into filamentous M13mp8 and M13WB2348 phages. Bioorg Khim. 1984;10(11):1544-7.
[2]
Paton EB, Vudmaska MI, Sverdlov ED. Unidirectional orientation of the rpoB gene of E. coli, cloned in filamentous phage Ml3. Macromolecules in the functioning cell: Proc. of reports IV sympos. USSR - Italy. Kiev, 1984; 89.
[3]
Paton EB, Woodmaska MI, Sverdlov ED. The stability of recombinant filamentous phages is reduced in the presence of the rpoB gene promoter. Biopolym Cell. 1985; 1(3):160-2.
[4]
Gentz R, Langner A, Chang AC, Cohen SN, Bujard H. Cloning and analysis of strong promoters is made possible by the downstream placement of a RNA termination signal. Proc Natl Acad Sci U S A. 1981;78(8):4936-40.
[5]
Ovchinnikov YA, Guryev SO, Krayev AS, Monastyrskaya GS, Skryabin KG, Sverdlov ED, Zakharyev VM, Bayev AA. Primary structure of an EcoRI fragment of lambda imm434 DNA containing regions cI-cro of phage 434 and cII-o of phage lambda. Gene. 1979;6(3):235-49.
[6]
Dretzen G, Bellard M, Sassone-Corsi P, Chambon P. A reliable method for the recovery of DNA fragments from agarose and acrylamide gels. Anal Biochem. 1981;112(2):295-8.
[7]
Maniatis T., Fritsch E. F., Sambrook J. Molecular cloning - a laboratory manual. New York : Cold Spring Harbor, 1982. 545 p.
[8]
Miller JH. Experiments in molecular genetics. Cold Spring Harbor Laboratory, 1972, 466 p.
[9]
Beck E, Zink B. Nucleotide sequence and genome organisation of filamentous bacteriophages fl and fd. Gene. 1981;16(1-3):35-58.
[10]
Collins J. Deletions, insertions and rearrangements affecting rpoB gene expression. Mol Gen Genet. 1979;173(2):217-20.