Biopolym. Cell. 1988; 4(3):163-167.
Short Communications
Orientational effects caused in the pUC plasmid by E. coli rplJL-rpoBC-operon fragments
1Paton E. B., 1Kroupskaya I. V., 1Zhyvoloup A. N.
  1. Institute of Molecular Biology and Genetics, Academy of Sciences of the Ukrainian SSR
    Kiev, USSR


A number of recombinant pUC plasmids containing E. coli rplJL-rpoBC-operon fragments were constructed. A rplJ gene and PL10 promoter containing 1568 bp EcoRI fragment and rplJ gene structural part containing BspRI-A-fragment were found to be oriented in pUC plasmids unidirectionally, in such a way that genes rplJ of the inserted fragment and lacZ' of the vector were transcribed in the opposite direction.


[1] Close TJ, Christmann JL, Rodriguez RL. M13 bacteriophage and pUC plasmids containing DNA inserts but still capable of beta-galactosidase alpha-complementation. Gene. 1983;23(2):131-6.
[2] Davanloo P, Rosenberg AH, Dunn JJ, Studier FW. Cloning and expression of the gene for bacteriophage T7 RNA polymerase. Proc Natl Acad Sci U S A. 1984;81(7):2035-9.
[3] Barry G, Squires CL, Squires C. Control features within the rplJL-rpoBC transcription unit of Escherichia coli. Proc Natl Acad Sci U S A. 1979;76(10):4922-6.
[4] Paton EB, Zhyvoloup AN, Varanitsa LA. The presence of two strong promoters determines the orientation of a DNA fragment inserted into pUC19 plasmid. Biopolym. Cell. 1986; 2(4):217-9.
[5] Paton EB., Kroupskaya IV, Zhyvoloup AN. Particularities of cloning of Escherichia coli rpoBC-operon fragments in the pUC plasmids. Biopolym. Cell. 1987; 3(6):307-12.
[6] 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.
[7] Fiil NP, Friesen JD, Downing WL, Dennis PP. Post-transcriptional regulatory mutants in a ribosomal protein-RNA polymerase operon of E. coli. Cell. 1980;19(4):837-44.
[8] Friesen JD, An G, Fiil N. The lethal effect of a plasmid resulting from transcriptional readthrough of rplJ from the rplKA operon in Escherichia coli. Mol Gen Genet. 1983;189(2):275-81.
[9] Zinder ND, Boeke JD. The filamentous phage (Ff) as vectors for recombinant DNA--a review. Gene. 1982;19(1):1-10.
[10] Miller JH. Experiments in molecular genetics. New York, Cold Spring Harbor Lab. press, 1972; 466 p.
[11] Maniatis T., Fritsch E. F., Sambrook J. Molecular cloning - a laboratory manual. New York: Cold Spring Harbor Lab., 1982. 545 p.
[12] Birnboim HC, Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979;7(6):1513-23.
[13] Clewell DB, Helinski DR. Supercoiled circular DNA-protein complex in Escherichia coli: purification and induced conversion to an opern circular DNA form. Proc Natl Acad Sci U S A. 1969;62(4):1159-66.
[14] Woodmaska MI, Paton EB. Cloning of Escherichia coli rpl JL-rpoBC-operon fragment in pBR322 and pHSG415 plasmids. Doklady Akad Nauk Ukr SSR. Ser B. 1987; (9):58-60.
[15] Downing WL, Dennis PP. Transcription products from the rplKAJL-rpoBC gene cluster. J Mol Biol. 1987;194(4):609-20.
[16] Post LE, Strycharz GD, Nomura M, Lewis H, Dennis PP. Nucleotide sequence of the ribosomal protein gene cluster adjacent to the gene for RNA polymerase subunit beta in Escherichia coli. Proc Natl Acad Sci U S A. 1979;76(4):1697-701.
[17] Stueber D, Bujard H. Transcription from efficient promoters can interfere with plasmid replication and diminish expression of plasmid specified genes. EMBO J. 1982;1(11):1399-404.
[18] Stassi DL, Lacks SA. Effect of strong promoters on the cloning in Escherichia coli of DNA fragments from Streptococcus pneumoniae. Gene. 1982;18(3):319-28.
[19] 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.
[20] Yanisch-Perron C, Vieira J, Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103-19.