Details

Autor(en) / Beteiligte

• Bai, Li-Li
• Yin, Wei-Bo
• Chen, Yu-Hong
• Niu, Li-Li
• Sun, Yong-Ru
• Zhao, Shi-Min
• Yang, Fu-Quan
• Wang, Richard R.-C
• Wu, Qing
• Zhang, Xiang-Qi
• Hu, Zan-Min
• Isalan, Mark

Titel

New Strategy to Produce a Defensin: Stable Production of Mutated NP-1 in Nitrate Reductase-Deficient Chlorella ellipsoidea

Ist Teil von

• PloS one, 2013-01, Vol.8 (1), p.e54966-e54966

Ort / Verlag

United States: Public Library of Science

Erscheinungsjahr

2013

Quelle

MEDLINE

Beschreibungen/Notizen

• 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 Npt II 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.