The HSP70A promoter as a tool for the improved expression of transgenes in Chlamydomonas.
The Chlamydomonas reinhardtii HSP70A promoter can be induced by both heat shock and light. Several characteristics of this promoter suggest its usefulness as a tool for improved transgene expression in this alga. (i) It may by itself confer high inducibility to a transgene. Fusion of the HSP70A promoter to reporter genes HSP70B or ARS yields high levels of transgene product that, as shown for ARS, may accumulate when repeated cycles of heat shock induction are applied. (ii) It activates other promoters. Using HSP70B as a reporter gene, we show that the HSP70A promoter serves as a transcriptional activator when placed upstream of the promoters RBCS2, beta 2 TUB and HSP70B. Activation of these promoters was observed both under basal conditions and upon light induction. In addition, transformation rates obtained for the eubacterial resistance gene aadA were significantly increased, when expression of this gene was controlled by the HSP70A-RBCS2 promoter fusion as compared to the RBCS2 promoter alone.
Schroda
M
M
Institute for Biology III, University of Freiburg, Germany.
Blöcker
D
D
Beck
C F
CF
D000818:Animals; D015152:Blotting, Northern; D016825:Chlamydomonas reinhardtii / Q000235:genetics* / Q000378:metabolism; D018506:Gene Expression Regulation, Plant; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics* / Q000378:metabolism; D006358:Hot Temperature; D008027:Light; D010940:Plant Proteins; D011401:Promoter Regions, Genetic; D015800:Protozoan Proteins; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014170:Transformation, Genetic
D018840:HSP70 Heat-Shock Proteins; C102156:HSP70B protein, Chlamydomonas; D010940:Plant Proteins; D015800:Protozoan Proteins; D011993:Recombinant Fusion Proteins