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Expression of the gene coding for a thermostable alpha-amylase from Pyrococcus furious in Chiamydomonas reinhardtii chloroplast / 生物工程学报
Chinese Journal of Biotechnology ; (12): 545-549, 2006.
Article in Chinese | WPRIM | ID: wpr-286252
Responsible library: WPRO
ABSTRACT
Thermostable alpha-amylase from Pyrococcus furious is an important industrial enzyme in brewing and alcohol production. Eexpression of the thermostable a-amylase in plants can reduce greatly costs in the production of alcohol using crop plants. A chloroplast expression vector, p64A, containing the thermostable alpha-amylase gene from Pyrococcus furious, was constructed with clpP-trnL-petB-chlL-rp123-rpl2 as Chlamydomonas reinhardtii plastid homologous recombinant fragments and spetinomycin-resistant aadA gene as select marker. The plasmid p64A was transferred into the chloroplast genome of C. reinhardtii by the biolistic method. Nine independently transformed lines were obtained by 100 mg/L spectinomycin selection. PCR amplification, Southern blot analysis of the transgene and cultivation in the dark all showed that the a-amylase gene had been integrated into chloroplast genome of C. reinhardtii. The activity of amylase expressed in the chloroplast of C. reinhardtii was detected by amylase activity assay and found to be as high as 77.5 u/g fresh weight of cells. These experimental results demonstrated the possibility of using transgenic chloroplasts of plant as bioreactors for production of industrial enzymes.
Subject(s)
Full text: Available Index: WPRIM (Western Pacific) Main subject: Plasmids / Enzyme Stability / Chemistry / Chloroplasts / Polymerase Chain Reaction / Chlamydomonas reinhardtii / Pyrococcus furiosus / Alpha-Amylases / Genetics / Metabolism Language: Chinese Journal: Chinese Journal of Biotechnology Year: 2006 Type: Article

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Full text: Available Index: WPRIM (Western Pacific) Main subject: Plasmids / Enzyme Stability / Chemistry / Chloroplasts / Polymerase Chain Reaction / Chlamydomonas reinhardtii / Pyrococcus furiosus / Alpha-Amylases / Genetics / Metabolism Language: Chinese Journal: Chinese Journal of Biotechnology Year: 2006 Type: Article