Expression Oleosin-MT Fusion Protein From Arabidopsis Thaliana And Safflower

MT (metallothionein) is a multi-functional induced protein, which exists in almost allorganisms (the molecular weight of6~8kDa). MT has anti-radiation, anti-oxidation, scavengingfree radicals and other reactive, it has been added to cosmetics widely, become a new cosmeticmaterials.Using plant system produces medicinal protein is one of the hot topics research in our time.Use plant oil-body expression heterologous proteins, not only reduce the production cost,simplify the extraction process, and lay a foundation for large-scale production of heterologousproteins.This study used plant oil system express Oleosin-MT, the antioxidant function ofmetallothionein is verified. By querying the base sequence of metallothionein inGenbank(NO.U11256.1),design specific primers，PCR amplification clone metallothionein gene,and with the oil body protein fusion，become an Oleosin-MT fusion protein. The fusion geneinserted into a p1301expression vector. Plant expression vectors p1301-OP-oleosin-MT wasconstructed, floral dip method infection Arabidopsis (Colombia.) and safflowers(Carthamustinctorius L.)，and the following conclusions is the result of Arabidopsis and safflowerexpression of Oleosin-MT:1.Detection the transgenic Arabidopsis plants in the molecular biology, fusion protein of26kDa was expressed in Arabidopsis seeds by verified SDS-PAGE and a good antioxidantactivity was detected through experiment in vitro.2.Oleosin-MT were successfully expressed in Arabidopsis，and it is suggested thatrecombinant protein had antioxidant activity.3. The Oleosin-MT gene has been integrated into the safflower genome.This study short the time safflower tissue culture,optimized the Basta quantity and time，and select the resistance safflower plants more efficient. This is the first trial of MT productionin an oleosin expression system，and the recombinant protein exhibited activity. This experimentconfirmed that the oleosin system express heterologous protein，provide a theoretical basis andlaid a good foundation for the health care products and cosmetics.