Biopolym. Cell. 1987; 3(5):263-269.
Gene-Engineering Biotechnology
Vectors for cloning and activity determination of the elements of the bacterial transcription system: cloning the transcription terminator of the rRNA operon of the archaebacterium Halobacterium halobium
1Skripkin E. A., 1Mankin A. S., 1Kopylov A. M., 2Khudyakov Yu. E.
  1. M. V. Lomonosov State University
    Moscow, USSR
  2. D. I. Ivanovsky Institute of Virology, Academy of Medical Sciences of the USSR
    Moscow, USSR

Abstract

The pBR322 vector was used for constructing recombinant plasmids pMM24 and pVR41, containing the promoter region PR of the bacteriophage λ including the transcription and translation starting points of the cro-repressor, the polylinker with the restriction sites for several restrictases and the tandem of the terminators of the fd phage. The constructed mini-gene is transcribed in E. coli cells into the corresponding mini-mRNA. The transcription of this gene could occur either constitutively or under the conduction of the c1857 represser of the phage λ. The polylinker sequence in the gene body could be used to introduce various regulator structures between the promoter and the terminators. A putative transcription terminator of the rRNA operon from the archaebacterium H. halobium was cloned inside the mini-gene. This archaebacterial sequence was shown to function as an effective terminator in the eubacterial system.

References

[1] Zabeau M, Stanley KK. Enhanced expression of cro-beta-galactosidase fusion proteins under the control of the PR promoter of bacteriophage lambda. EMBO J. 1982;1(10):1217-24.
[2] Maniatis T., Fritsch E. F., Sambrook J. Molecular cloning: a laboratory manual New York: Cold Spring Harbor Lab. publ., 1982 545 p.
[3] Mukhopadhyay M, Mandal NC. A simple procedure for large-scale preparation of pure plasmid DNA free from chromosomal DNA from bacteria. Anal Biochem. 1983;133(2):265-70.
[4] Maxam AM, Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499-560.
[5] Jinks-Robertson S, Gourse RL, Nomura M. Expression of rRNA and tRNA genes in Escherichia coli: evidence for feedback regulation by products of rRNA operons. Cell. 1983;33(3):865-76.
[6] von Gabain A, Belasco JG, Schottel JL, Chang AC, Cohen SN. Decay of mRNA in Escherichia coli: investigation of the fate of specific segments of transcripts. Proc Natl Acad Sci U S A. 1983;80(3):653-7.
[7] Ikemura T, Itoh S, Post LE, Nomura M. Isolation and characterization of stable hybrid mRNA molecules transcribed from ribosomal protein promoters in E. coli. Cell. 1979;18(3):895-903.
[8] Hui I, Dennis PP. Characterization of the ribosomal RNA gene clusters in Halobacterium cutirubrum. J Biol Chem. 1985;260(2):899-906.
[9] Daniels CJ, Hofman JD, MacWilliam JG, Doolittle WF, Woese CR, Luehrsen KR, Fox GE. Sequence of 5S ribosomal RNA gene regions and their products in the archaebacterium Halobacterium volcanii. Mol Gen Genet. 1985;198(2):270-4.
[10] Moritz A, Lankat-Buttgereit B, Gross HJ, Goebel W. Common structural features of the genes for two stable RNAs from Halobacterium halobium. Nucleic Acids Res. 1985;13(1):31-43.
[11] Mankin AS, Kagramanova VK. Complete nucleotide sequence of the single ribosomal RNA operon of Halobacterium halobium: secondary structure of the archaebacterial 23S rRNA. Mol Gen Genet. 1986; 202(1):152-161.
[12] Donis-Keller H, Maxam AM, Gilbert W. Mapping adenines, guanines, and pyrimidines in RNA. Nucleic Acids Res. 1977;4(8):2527-38.