nickel and low co2-controlled motility in chlamydomonas through complementation of a paralyzed flagella mutant with chemically regulated promoters镍和低co2-controlled能动性在衣藻通过互补的瘫痪鞭毛与化学调控启动子突变.pdfVIP

Ferrante et al. BMC Plant Biology 2011, 11:22 /1471-2229/11/22 RESEARCH ARTICLE Open Access Nickel and low CO -controlled motility in 2 Chlamydomonas through complementation of a paralyzed flagella mutant with chemically regulated promoters 1 2 2 1* Paola Ferrante , Dennis R Diener , Joel L Rosenbaum , Giovanni Giuliano Abstract Background: Chlamydomonas reinhardtii is a model system for the biology of unicellular green algae. Chemically regulated promoters, such as the nickel-inducible CYC6 or the low CO2-inducible CAH1 promoter, may prove useful for expressing, at precise times during its cell cycle, proteins with relevant biological functions, or complementing mutants in genes encoding such proteins. To this date, this has not been reported for the above promoters. Results: We fused the CYC6 and CAH1 promoters to an HA-tagged RSP3 gene, encoding a protein of the flagellar radial spoke complex. The constructs were used for chemically regulated complementation of the pf14 mutant, carrying an ochre mutation in the RSP3 gene. 7 to 8% of the transformants showed cells with restored motility after induction with nickel or transfer to low CO2 conditions, but not in non-inducing conditions. Maximum complementation (5% motile cells) was reached with very different kinetics (5-6 hours for CAH1, 48 hours for CYC6). The two inducible promoters drive much lower levels of RSP3 protein expression than the constitutive PSAD promoter, which shows almost complete rescue of motility. Conclusions: To our knowledge, this is the first example of the use of the CYC6 or CAH1 promoters to perform a chemically regulated complementation of a Chlamydomonas mutant. Based on our data, the CYC6 and CAH1 promoter