Isolation and characterization of a nitrate reductase deficient mutant of Chlorella ellipsoidea (Chlorophyta)

Wang, Yiqin; Zhang, Xiaoyu; Peng, Wang; Geng, Deqin; Zhao, Shimin; Zhang, Liming; Sun, Yuhan

doi:10.1007/s10811-005-5559-7

Cited by 12 publications

(10 citation statements)

References 16 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Therefore, NR gene can be used as a selective marker when nrm -4 was used as the host cell using nitrate as the selection chemical. NR gene from C. ellipsoidea was cloned and characterized by our group [27]. Our previous work also proved that Npt II gene could be used as a selective marker in the transformation of C. ellipsoidea using G418 as the selection chemical [26].…”

Section: Discussionmentioning

confidence: 95%

“…NR-deficient C. ellipsoidea mutant nrm -4 was previously developed by our group using X-ray mutagenesis method [27]. It grew in the medium with nitrite,but not in the medium with nitrate.…”

Section: Discussionmentioning

confidence: 99%

“…NR-deficient C. ellipsoidea mutant nrm -4 [27] was used as the host cells. The stock culture of nrm -4 was maintained on an E4 agar slant medium containing SE (Brostol’s solution) [28] with modification, in which NaNO 3 was substituted with 0.7 g/l NaNO 2 and 15 g/l agar.…”

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

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

et al. 2013

View full text Add to dashboard Cite

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 NaNO3. 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.

show abstract

Section: Discussionmentioning

confidence: 95%

“…NR-deficient C. ellipsoidea mutant nrm -4 was previously developed by our group using X-ray mutagenesis method [27]. It grew in the medium with nitrite,but not in the medium with nitrate.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

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

et al. 2013

View full text Add to dashboard Cite

show abstract

“…Nuclear transformation systems have been developed by introducing the NR gene (NIA1) as a selectable marker into NR-deficient mutants in Chlamydomonas (Kindle et al, 1989), Volvox (Schiedlmeier et al, 1994), and Chlorella (Dawson et al, 1997;Wang et al, 2005;Zhang et al, 2005). However, whether this selection system can be used for transgenic D. salina has yet to be determined.…”

Section: Introductionmentioning

confidence: 99%

“…Because NR mutants with defects in the nitrogen assimilation pathway display chlorate resistance, screening for chlorate sensitivity has been successfully used for isolating mutants from several kinds of algae, such as Chlorella ellipsoidea (Wang et al, 2005) and Dunaliella viridis (Sun et al, 2006). We have previously demonstrated that wild-type D. salina can not grow in liquid media containing 200 mM chlorate or on culture plates containing 100 mM chlorate (Lu et al, 2011).…”

Section: Introductionmentioning

confidence: 99%

Characterization of novel nitrate reductase-deficient mutants for transgenic Dunaliella salina systems

Lj¹,

Jia²,

Sk³

et al. 2015

Genet. Mol. Res.

View full text Add to dashboard Cite

ABSTRACT. The aim of the present study was to isolate and characterize novel nitrate reductase (NR)-deficient mutants, which may be useful for the transgenic manipulation of Dunaliella salina. Three NR-deficient mutants of D. salina, J-1, J-2, and J-3, were successfully isolated by screening for chlorate resistance after chemical mutagenesis with ethylnitrosourea. NR activity was not detected in the mutants and the expression of NR mRNA was significantly decreased. Growth analysis of D. salina strains grown in media containing different nitrogen sources revealed that these mutants were capable of utilizing nitrite and urea, but not nitrate as a nitrogen source, indicating that these mutants are indeed NR-deficient. Mutation analysis of NR cDNA sequences revealed that there were 11 point mutations shared by the J-1, J-2, and J-3 mutants. Furthermore, the results of the functional complementation experiment showed that NR activity of transformant T-1 derived from J-1 was recovered to 48.1 % of that of the wild-type D. salina. The findings of the present study indicate that nitrate may be used as a selective agent rather than antibiotics or herbicides for the isolated NRdeficient mutants in future transgenic D. salina systems.

show abstract

Biology and Industrial Applications of Chlorella: Advances and Prospects

Liu

Chen

2014

Advances in Biochemical Engineering/Biotechnology

View full text Add to dashboard Cite

Chlorella represents a group of eukaryotic green microalgae that has been receiving increasing scientific and commercial interest. It possesses high photosynthetic ability and is capable of growing robustly under mixotrophic and heterotrophic conditions as well. Chlorella has long been considered as a source of protein and is now industrially produced for human food and animal feed. Chlorella is also rich in oil, an ideal feedstock for biofuels. The exploration of biofuel production by Chlorella is underway. Chlorella has the ability to fix carbon dioxide efficiently and to remove nutrients of nitrogen and phosphorous, making it a good candidate for greenhouse gas biomitigation and wastewater bioremediation. In addition, Chlorella shows potential as an alternative expression host for recombinant protein production, though challenges remain to be addressed. Currently, omics analyses of certain Chlorella strains are being performed, which will help to unravel the biological implications of Chlorella and facilitate the future exploration of industrial applications.

show abstract

Isolation and characterization of a nitrate reductase deficient mutant of Chlorella ellipsoidea (Chlorophyta)

Cited by 12 publications

References 16 publications

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

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

Characterization of novel nitrate reductase-deficient mutants for transgenic Dunaliella salina systems

Biology and Industrial Applications of Chlorella: Advances and Prospects

Contact Info

Product

Resources

About