Highly efficient expression of rabbit neutrophil peptide-1 gene in Chlorella ellipsoidea cells

Chen, Ying; Wang, Yiqin; Sun, Yuhan; Zhang, Liming; Li, Wenbin

doi:10.1007/s002940100205

Cited by 82 publications

(53 citation statements)

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“…has been hindered by the lack of genomics research models. Moreover, although genetic modification of Chlorella vulgaris (Chow and Tung, 1997) and Chlorella ellipsoidea (Chen et al, 2001) were achieved, none of the Chlorella spp. strains shown to be amenable to transformation are of commercial interest (Tran et al, 2013).…”

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confidence: 99%

Genomic Foundation of Starch-to-Lipid Switch in Oleaginous Chlorella spp.

et al. 2015

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The ability to rapidly switch the intracellular energy storage form from starch to lipids is an advantageous trait for microalgae feedstock. To probe this mechanism, we sequenced the 56.8-Mbp genome of Chlorella pyrenoidosa FACHB-9, an industrial production strain for protein, starch, and lipids. The genome exhibits positive selection and gene family expansion in lipid and carbohydrate metabolism and genes related to cell cycle and stress response. Moreover, 10 lipid metabolism genes might be originated from bacteria via horizontal gene transfer. Transcriptomic dynamics tracked via messenger RNA sequencing over six time points during metabolic switch from starch-rich heterotrophy to lipid-rich photoautotrophy revealed that under heterotrophy, genes most strongly expressed were from the tricarboxylic acid cycle, respiratory chain, oxidative phosphorylation, gluconeogenesis, glyoxylate cycle, and amino acid metabolisms, whereas those most down-regulated were from fatty acid and oxidative pentose phosphate metabolism. The shift from heterotrophy into photoautotrophy highlights up-regulation of genes from carbon fixation, photosynthesis, fatty acid biosynthesis, the oxidative pentose phosphate pathway, and starch catabolism, which resulted in a marked redirection of metabolism, where the primary carbon source of glycine is no longer supplied to cell building blocks by the tricarboxylic acid cycle and gluconeogenesis, whereas carbon skeletons from photosynthesis and starch degradation may be directly channeled into fatty acid and protein biosynthesis. By establishing the first genetic transformation in industrial oleaginous C. pyrenoidosa, we further showed that overexpression of an NAD(H) kinase from Arabidopsis (Arabidopsis thaliana) increased cellular lipid content by 110.4%, yet without reducing growth rate. These findings provide a foundation for exploiting the metabolic switch in microalgae for improved photosynthetic production of food and fuels.

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confidence: 99%

Genomic Foundation of Starch-to-Lipid Switch in Oleaginous Chlorella spp.

et al. 2015

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“…More recently, Geng et al (2003) described both the stable and transient nuclear transformation of D. salina using electroporation and expression of the hepatitis B surface antigen under the control of the maize ubiquitin-promoter, a promoter that has previously been used for the expression of foreign genes in Chlorella ellipsoidea (Chen et al, 2001). While the rate of transient transformation was relatively low, at least 14 positive lines were confirmed by PCR and Southern blot analysis although stable integration of the transgene was not determined.…”

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confidence: 96%

Towards the development of a nuclear transformation system for Dunaliella tertiolecta

et al. 2005

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A nuclear transformation system for the microalga Dunaliella tertiolecta was explored using electroporation. Plasmids incorporating the D. tertiolecta RbcS1 5 and 3 untranslated regions flanking the Streptoalloteichus hindustanus gene encoding bleomycin resistance (ble) were introduced into D. tertiolecta cells both transiently and stably. Southern hybridisation was used to examine the fate of the plasmid following electroporation and revealed that the DNA was entering the cells but was quickly degraded. Using the same procedure one stably transformed line was recovered.Abbreviations: ble, bleomycin resistance gene; RbcS, ribulose bisphosphate carboxylase/oxygenase small subunit

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“…Results showed that the transformed plants inhibited B. subtilis and P. aeruginosa growth, which was consistent with the results of NP-1 genes introduced into poplar and Chlorella (Zhao et al, 1999;Wang et al, 2001). The NP-1 disease-resistant mechanism was combined with a virus coat protein to lose their biological activity (Chen et al, 2001b). However, only three strains showed better inhibitory effects in six positive transgenic plants, which may be owing to the following: first, the low expression of the NP-1 gene; there is a threshold level of transgene-derived transcripts, and disease-resistant mechanisms cannot be triggered until the threshold level in transgenic plants is reached (Fu et al, 1998); second, the genetic variation of other disease-resistant related genes during the process of transformation; and third, false positives.…”

Section: Discussionmentioning

confidence: 99%

Transfer and expression of the rabbit defensin NP-1 gene in lettuce (Lactuca sativa)

Song

Xiong

et al. 2017

Genet. Mol. Res.

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ABSTRACT. Lettuce (Lactuca sativa L.) is an annual plant of the daisy family, Asteraceae, with high food and medicinal value. However, the crop is susceptible to several viruses that are transmitted by aphids and is highly vulnerable to post-harvest diseases, as well as insect and mammal pests and fungal and bacterial diseases. Here, the rabbit defensin gene NP-1 was transferred into lettuce by Agrobacteriummediated transformation to obtain a broad-spectrum disease-resistant lettuce. Transgenic lettuce plants were selected and regenerated on selective media. The presence of the NP-1 gene in these plants was confirmed by western blot analyses. Resistance tests revealed native defensin NP-1 expression conferred partial resistance to Bacillus subtilis and Pseudomonas aeruginosa, which suggests new possibilities for lettuce disease resistance.

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Highly efficient expression of rabbit neutrophil peptide-1 gene in Chlorella ellipsoidea cells

Cited by 82 publications

References 0 publications

Genomic Foundation of Starch-to-Lipid Switch in Oleaginous Chlorella spp.

Genomic Foundation of Starch-to-Lipid Switch in Oleaginous Chlorella spp.

Towards the development of a nuclear transformation system for Dunaliella tertiolecta

Transfer and expression of the rabbit defensin NP-1 gene in lettuce (Lactuca sativa)

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