Regulated Expression of lacZ Gene Fusions in Tobacco
Analysis of gene regulation with fusions to reporter genes has proved to be successful in many biological systems including both prokaryotes and eukaryotes. The regulatory regions of the genes under study are joined to the coding sequence of the reporter gene and subsequent to transfer of the gene fusion into the host cell, the activity of the reporter gene product will reflect the activity of the regulatory regions. A good choice of the reporter gene allows convenient, standard and quantitative meas?rement of gene expression and, in some cases, detection of reporter gene activity directly in the tissue. Genes such as nptll, cat, luc and uidA have been used to monitor the activity of plant promoters controlling their expression in transgenic plants (Bevan et al. 1983 ; Herrera-Estrella et al. 1983 ; Ow et al. 1986; Jefferson et al. 1987). These genes code for the enzymes neomycin phosphotransferase II, chloramphenicol acetyltransferase, the firefly luciferase and β-glucuronidase (GUS) respectively, which are relatively easy to assay.
KeywordsReporter Gene Gene Fusion Galactosidase Activity Chloramphenicol Acetyltransferase LacZ Activity
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- Bassford P, Beckwith J, Berman M, Brickman E, Casadaban M, Guarente L, Saint-Girons I, Sarthy A, Schwartz M, Shuman H, Silhavy T (1978) Genetic fusions of the LAC Operon: a new approach to the study of biological processes. In: Miller JH, Reznikoff WS (eds) The Operon. Cold Spring Harbor Press, New York, pp 245–261Google Scholar
- Cashmore AR (1983) Nuclear genes encoding the small subunit of ribulose-l,5-bisphosphate carboxylase. In: Kosuge T, Meredith CP, Hollaender A (eds) Genetic engineering of plants. Plenum, New York, pp 29–38Google Scholar
- De Greve H, Dhaese P, Seurink J, Lemmers M, Van Montagu M, Schell J (1982) Nucleotide sequence and transcript map of the Agrobacterium tumefaciens Ti plasmid encoded octopine synthase gene. J Mol Appl Gen 1:499–511Google Scholar
- Depicker A, Stachel S, Dhaese P, Zambryski P, Goodman HM (1982) Nopaline synthase transcript mapping and DNA sequence. J Mol Appl Gen 1:561–573Google Scholar
- Herrer-Estrella L, De Block M, Messens E, Hernalsteens J-P, Van Montagu M, Schell J (1983) Chimeric genes as dominant selectable markers in plant cells. EMBO J 2:987–995Google Scholar
- Kamins A, Otto K, Rüther U, Müller-Hill B (1983) Sequence of the lacZ gene of Escherichia coli. EMBO J 2:593–597Google Scholar
- Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor Lab Press, New YorkGoogle Scholar
- UUman A, Perrin D (1970) Complementation in beta-galactosidase. In: Beckwith JR, Zipser D (eds) The lactose Operon. Cold Spring Harbor Lab Press, New York, pp 143–172Google Scholar