The function of plant heat shock promoter elements in the regulated expression of chimaeric genes in transgenic tobacco.
A series of deletion mutants of a soybean heat shock (hs) gene promoter was generated and linked to the chloramphenicol acetyl transferase (CAT) coding sequence. These chimaeric promoter/reporter gene constructs were introduced into tobacco and thermoregulated expression of CAT activity was examined in leaf extracts. Three different types of gene fusions were tested using two different BIN19 vector constructions: (1) translational fusion between the N-terminus of the protein coding sequence of the heat shock gene Gmhsp17.3-B and CAT; (2) transcriptional fusions between the 5' nontranslated RNA regions of Gmhsp17.3-B and CAT; and (3) promoter fusions joining the hs promoter upstream sequences to the TATA box sequence of the delta CaMV 35S-CATter vector. Alternatively, multiple copies of a synthetic deoxyoligonucleotide with the soybean hs consensus element (HSE2) were used. Heat inducible CAT activities were detected except in plants containing a transcriptional fusion devoid of all but 18 nucleotides at the 5' terminus of the hs gene transcript. CAT activity was detectable in these plants only during the recovery at 25 degrees C after a hs (40 degrees C). Overlapping HSE-like promoter sequences seem to be necessary for the induction of heat inducible transcription of linked genes; synthetic HSE2 sequences have the capacity to reconstitute a hs promoter in combination with a TATA box sequence. Effective translation during hs seems to require sequences in the 5' non-translated leader of the hs protein mRNA; these sequences can be functionally replaced by the 5' leader sequence of the delta CaMV 35S promoter.
Universität Bielefeld, Fakultät für Biologie (Genetik), Federal Republic of Germany.
F
F
Schöffl
M
M
Rieping
G
G
Baumann
M
M
Bevan
S
S
Angermüller
D000595:Amino Acid Sequence; D001483:Base Sequence; D002678:Chimera; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D002872:Chromosome Deletion; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D005786:Gene Expression Regulation; D006360:Heat-Shock Proteins / Q000235:genetics*; D008969:Molecular Sequence Data; D010944:Plants / Q000235:genetics*; D010947:Plants, Toxic; D011401:Promoter Regions, Genetic; D014026:Nicotiana / Q000235:genetics; D014170:Transformation, Genetic
D006360:Heat-Shock Proteins; D004247:DNA; D015500:Chloramphenicol O-Acetyltransferase