Summary
This study presents plasmid systems that utilize regulatory signals of bacteriophage Lambda to accomplish regulated expression of cloned genes in an A. nidulans R2 derivative strain. An operator-promoter region and the temperature-sensitive repressor gene cI857 of bacteriophage Lambda were employed. Linked to a cyanobacterial replicon, the plasmid vectors efficiently transformed Anacystis and were stably maintained within this host. The cat structural gene, encoding chloramphenicol acetyltransferase, was used to demonstrate that expression can be regulated by temperature shift. We have identified in extracts from the vector bearing Anacystis, a protein similar in size and immunology to the Lambda repressor. The systems described should allow controlled expression of adventitious genes in the cyanobacterial host.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- APr :
-
ampicillin resistance
- Cmr :
-
chloramphenicol resistance
- CmActase:
-
chloramphenicol acetyltransferase
- Kmr :
-
Kanamycine resistance
- [ ]:
-
indicates plasmid carrier state
References
Close TJ, Rodriguez RL (1982) Construction and characterization of the chloramphenicol-resistance gene cartridge: a new approach to the transcriptional mapping of extrachromosomal elements. Gene 20:305–316
Covarrubias L, Bolivar F (1982) Construction and characterization of new cloning vehicles. VI. plasmid pBR329, a new derivative of pBR328 lacking the 482-base-pare inverted duplication. Gene 17:79–89
Edelman M, Reisfeld (1980) Synthesis, processing and functional probing of P-32000, the major membrane protein translated within the chloroplast. In: Leaver CJ (ed) Genome Organization and expression in plants, Plenum Press, New York pp 353–362
Friedberg D, Goldfarb D (1983) Expression of chloramphenicol resistance in Escherichia coli by Anacystis nidulans 6311 promoters In: 15th FEBS meeting p 112
Friedberg D, Seijffers J (1979) Plasmids in two cyanobacterial strains. FEBS Lett 107:165–168
Friedberg D, Seijffers J (1983) A new hybrid plasmid capable of transforming Escherichia coli and Anacystis nidulans. Gene 22:267–275
Golden SS, Sherman LA (1983) A hybrid plasmid is a stable cloning vector for the cyanobacterium Anacystis nidulans R2. J Bacteriol 155:966–972
Gussin G, Jonson A, Pabo C, Sauer R (1983) Repressor and cro protein: structure, function and role in lysogenization. In: Hendrix RW, Roberts JW, Stahl FW, Weisberg RA (eds) Lambda II Cold Spring Harbor Laboratory pp 93–121
Herdman M (1982) Evolution and genetic properties of cyanobacterial genomes. In: Carr NG, Whitton BA (eds) The Biology of cyanobacteria. Blackwell, Oxford pp 263–305
Honigman A, Mahajna J, Altuvia S, Koby S, Teff D, Locker-Giladi H, Hayman H, Kronman C, Oppenheim AB (1985) Plasmid vectors for the analysis of transcription termination signals. Gene 36:131–141
Kolowsky KS, Williams GK, Szalay AA (1984) Length of foreign DNA in chimeric plasmids determines the efficiency of its integration into the chromosome of the cyanobacterium Synechococcus R2. Gene 27:289–299
Kuhlemeier CJ, Borrias WE, Van den Hondel CAMJJ, Van Arkel GA (1981) Vectors for cloning in cyanobacteria: construction and characterization of two recombinant plasmids capable of transformation to Escherichia coli K12 and Anacystis nidulans R2. Mol Gen Genet 184:249–254
Kuhlemeier CJ, Thomas AAM, Van der Ende A, Van Leen RW, Borrias WE, Van den Hondel CAMJJ, Van Arkel GA (1983) A host-vector system for gene cloning in the cyanobacterium Anacystis nidulans R2. Plasmid 10:156–163
Kuhlemeier CJ, Teeuwsen VJP, Janssen MJT, Van Arkel GA (1984) Cloning of a third nitrate reductase gene from the cyanobacterium Anacystis nidulans R2 using a shuttle cosmid library. Gene 31:109–116
Laban A, Cohen A (1981) Interplasmidic and intraplasmidic recombination in Escherichia coli. Mol Gen Genet 184:200–204
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
Lowry OH, Rosenbrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275
Maniatis T, Fritsch EF, Sambrook J (1982) 62) Seances Acad Sci 254:1517–1519
Miller SS, Ausubel FM, Bogorad L (1979) Cyanobacterial ribonucleic Acid polymerases recognise Lambda promoters. J Bacteriol 140:246–250
Oka A, Sugisaki H, Takanami M (1981) Nucleotide sequence of the kanamicine resistance transposon Tn903. J Mol Biol 147:217–226
Ptashne M, Backman K, Humayun MZ, Maurer R, Meyer B, Sauer RT (1976) Autoregulation and function of repressor in bacteriophage Lambda. Science 194:156
Rosenberg M, Court DA (1979) Regulatory sequences involved in the promotion and termination of RNA transcription. Ann Rev Genet 13:319–353
Rosenberg M, Ho Y, Shatzman A (1983) The use of pKC30 and its derivatives for controlled expression of genes. In: Wu R, Grossman L, Moldave K (eds) Methods in enzymology, Vol 101. Academic Press, New York, pp 123–138
Shaw WV (1983) Chloramphenicol acetyltransferase from chloramphenicol resistant bacteria. In: Hash JH (ed) Methods in enzymology, Vol 43. Academic Press, New York, pp 737–755
Shestakov SV, Khyen NT (1970) Evidence for genetic transformation in blue-green alga Anacystis nidulans. Mol Gen Genet 107:372–375
Shimatake H, Rosenberg M (1981) Purified lambda regulatory protein cII positively activates promoters for lysogenic development. Nature 292:128
Stanier RY, Cohen-Bazire G (1977) Phototrophic prokaryotes: the cyanobacteria. Ann Rev Microbiol 32:225–274
Sussman R, Jacob F, Hebel CR (1962) Sur un systeme de repression thermosensible chez le bacteriophage lambda d'Escherichia coli. CR Acad Sci 254:1517–1519
Tandeau de Marsac N, Borrias WE, Kuhlemeier CJ, Castets AM, Van Arkel GA, Van den Hondel CAMJJ (1982) A new approach for molecular cloning in cyanobacteria: cloning of an Anacystis nidulans met gene using Tn901-induced mutant. Gene 20:111–119
Tumer NE, Robinson SJ, Haselkorn R (1983) Different promoters for the Anabaena geutamine synthetase gene during growth using molecular or fixed nitrogen. Nature 306:337–342
Van den Hondel CAMJJ, Keegstra W, Borrias WE, Van Arkel GA (1979) Homology of plasmids in strains of unicellular cyanobacteria. Plasmid 2:323–333
Vieira J, Messing J (1982) The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primer. Gene 19:259–268
Williams JGK, Szalay AA (1983) Stable integration of foreign DNA into the chromosome of the cyanobacterium Synechococcus R2. Gene 24:37–51
Author information
Authors and Affiliations
Additional information
Communicated by M. Takanami
Rights and permissions
About this article
Cite this article
Friedberg, D., Seijffers, J. Controlled gene expression utilising Lambda phage regulatory signals in a cyanobacterium host. Mol Gen Genet 203, 505–510 (1986). https://doi.org/10.1007/BF00422077
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00422077