title: A new strategy to produce a defensin: stable production of mutated NP-1 in nitrate reductase-deficient Chlorella ellipsoidea.
abstract: Defensins are small cationic peptides that could be used as the potential substitute for antibiotics. However, there is no efficient method for producing defensins. In this study, we developed a new strategy to produce defensin in nitrate reductase (NR)-deficient C. ellipsoidea (nrm-4). We constructed a plant expression vector carrying mutated NP-1 gene (mNP-1), a mature α-defensin NP-1 gene from rabbit with an additional initiator codon in the 5'-terminus, in which the selection markers were NptII and NR genes. We transferred mNP-1 into nrm-4 using electroporation and obtained many transgenic lines with high efficiency under selection chemicals G418 and NaNO(3). The mNP-1 was characterized using N-terminal sequencing after being isolated from transgenic lines. Excitingly, mNP-1 was produced at high levels (approximately 11.42 mg/l) even after 15 generations of continuous fermentation. In addition, mNP-1 had strong activity against Escherichia coli at 5 µg/ml. This research developed a new method for producing defensins using genetic engineering.
authors: Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, People's Republic of China.
authors: Li-Li
authors: LL
authors: Bai
authors: Wei-Bo
authors: WB
authors: Yin
authors: Yu-Hong
authors: YH
authors: Chen
authors: Li-Li
authors: LL
authors: Niu
authors: Yong-Ru
authors: YR
authors: Sun
authors: Shi-Min
authors: SM
authors: Zhao
authors: Fu-Quan
authors: FQ
authors: Yang
authors: Richard R-C
authors: RR
authors: Wang
authors: Qing
authors: Q
authors: Wu
authors: Xiang-Qi
authors: XQ
authors: Zhang
authors: Zan-Min
authors: ZM
authors: Hu
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000096:biosynthesis* / Q000494:pharmacology; D002708:Chlorella / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism*; D018387:Codon, Initiator / Q000235:genetics; D004926:Escherichia coli / Q000187:drug effects; D005285:Fermentation; D005822:Genetic Vectors / Q000235:genetics; D009154:Mutation; D050901:Nitrate Reductase / Q000172:deficiency*; D030821:Plants, Genetically Modified; D011817:Rabbits; D014170:Transformation, Genetic; D023084:alpha-Defensins / Q000096:biosynthesis* / Q000235:genetics* / Q000494:pharmacology
chemical_list: D000900:Anti-Bacterial Agents; D018387:Codon, Initiator; D023084:alpha-Defensins; C406564:defensin NP-1; D050901:Nitrate Reductase
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