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| 1 | Simultaneously down-regulation of multiplex branch pathways using CRISPRi and fermentation optimization for enhancingβ-amyrin production in Saccharomyces cerevisiae显示文摘The production ofβ-amyrin in Saccharomyces cerevisiae is still low due to the inability of effectively regulating the endogenous metabolic pathway for competitive synthesis ofβ-amyrin precursors.In this study,we focused on two branches ofβ-amyrin synthetics pathway that consumeβ-amyrin precursors(2,3-oxidosqualene and cytosolic acetyl-CoA)and regulated related genes(ADH1,ADH4,ADH5,ADH6,CIT2,MLS2 and ERG7).We developed a CRISPRi method by constructing a multi-gRNA plasmid to down-regulate the seven genes simultaneously,which is reported for the first time in S.cerevisiae.The average transcription inhibition efficiency of the seven genes reached as high as 75.5%.Furthermore,by optimizing the fermentation condition(including pH,inoculum size,initial glucose concentration and feed of glucose or ethanol)and increasing extracellular transportation via supplying methyl-β-cyclodextrin,β-amyrin concentration of engineered strain SGibSdCg increased by 44.3%compared with the parent strain SGib,achieving 156.7 mg/L which was the highest concentration ofβ-amyrin reported in yeast.The one-step down-regulation of multiple genes using CRISPRi showed high efficiency and promising future in improving the yields of natural products. | Jiangping Ni Genlin Zhang Lei Qin Jun Li Chun Li | 2019 | Synthetic and Systems Biotechnology2019，4，2： | 4 |
| 2 | Genetic diversity of wild barley (Hordeum vulgare ssp. spontaneum) and its utilization for barley improvement显示文摘Wild barley(Hordeum vulgare spp.spontaneum) is the undisputed progenitor of cultivated barley and offers considerable potential as a genetic resource for barley improvement.Naturally occurring populations are distributed primarily across the Fertile Crescent,but secondary areas of diversity are also found in central Asia and Tibet.The pattern of its genetic diversity is non-random,and is structured on a global,regional and local scale.On a global scale,diversity follows ecogeography,while on a regional and local scale,it is more strongly determined by ecology.Populations growing in areas where the climate is unreliable and/or the soil is heterogeneous tend to retain the most diversity.A large number of accessions are currently conserved ex situ,but a high proportion of these have originated from the Fertile Crescent.Given the likelihood of future climate change,conserving germplasm both in situ and ex situ in the various regions where the species' currently existing is a research priority.The value of H.vulgare spp.spontaneum for the improvement of cultivated barley lies largely in its potential contribution to the levels of attainable pathogen resistance and abiotic stress tolerance. | XiaoYing Ma Chao Li AiDong Wang RuiJun Duan GenLin Jiao Eviatar Nevo GuoXiong Chen | 2012 | Research in Cold and Arid Regions2012，4，6： | 2 |
| 3 | Construction of a eukaryotic expression plasmid for human retina-derived neurotrophin-3显示文摘Neurotrophin-3 (NT-3) can promote the repair of central nervous system and retinal damage. In previous reports, NT-3 has been expressed by viral vectors. However, plasmid vectors have a safer profile compared with viral vectors in clinical studies. This study recombined amplified human retinal NT-3 with a eukaryotic expression plasmid containing green fluorescent protein (GFP) to construct an NT-3 expression plasmid, pEGFP-N1-NT-3. The transfection efficiency 48 hours after pEGFP-N1-NT-3 transfection to 293T cells was 50.06 ± 2.78%. Abundant NT-3-GFP was expressed in 293T cells as observed by fluorescence microscopy, suggesting the construct pEGFP-N1-NT-3 effectively expressed and secreted NT-3-GFP. Secretory vesicles containing NT-3-GFP were observed in a constant location in cells by laser scan confocal microscopy, indicating the expression and secretion processes of NT-3 in eukaryotic cells were in accordance with the physical synthesis processes of secreted proteins. Western blot assay showed that pro-NT-3-GFP had a molecular weight of 56 kDa, further confirming NT-3-GFP expression. At 48 hours after transfection, the concentration of NT-3 in culture medium was 22.3 ng/mL, suggesting NT-3 produced by pEGFP-N1-NT-3 was efficiently secreted. This study constructed a human retinal-derived NT-3 eukaryotic expression plasmid that efficiently expressed and secreted NT-3. | Chunxia Peng Xiaobei Yin Mengda Li Ting He Genlin Li | 2013 | Neural Regeneration Research2013，8，11： | 1 |
| 4 | Constitutive equation for elevated temperature flow behavior of TiNiNb alloy based on orthogonal analysis显示文摘 | Meili Xiao Fuguo Li Wei Zhao Genlin Yang | 2011 | Materials and Design2011，，： | 1 |
| 5 | Boosting the Rate Performance of Li–S Batteries via Highly Dispersed Cobalt Nanoparticles Embedded into Nitrogen-Doped Hierarchical Porous Carbon显示文摘Lithium–sulfur(Li–S)batteries are one of the most promising alternatives to lithium–ion batteries because of the advantageous high energy density and low cost.However,the practical applications of Li–S batteries are hampered by a severe shuttle effect and sluggish polysulfide redox conversion.Herein,highly dispersed cobalt nanoparticles(∼0.8 wt%)embedded into nitrogen-doped hierarchical porous carbon(Co@N-HPC)are designed as an effective electrocatalyst for Li–S batteries,which exhibit a synergistic effect of anchoring and dual-directional catalytic conversion of polysulfides. | Cheng Yuan Pan Zeng Chen Cheng Tianran Yan Genlin Liu Wenmin Wang Jun Hu Xin Li Junfa Zhu Liang Zhang | 2022 | CCS Chemistry2022，4，8： | 1 |
| 6 | Key Artificial Cultivation Technique by the Naxi Nationality for Dioscorea bulbifera:A Traditional Medicinal and Edible Plant显示文摘Dioscorea bulbifera is a medicinal and edible plant of the Naxi nationality living in Lijiang area,and its tuber as a vegetable has the functions of clearing away heat and detoxification,and its bulbil has the functions of clearing away heat and resolving phlegm,relieving cough and relieving asthma,dispersing the accumulation of evils and detoxification,etc.Related studies have shown that it mainly contains steroids,diterpene lactones,flavonoids,polyphenols,alkaloids and so on,and it can also fight cancer,inhibit bacteria,relieve pain,resist inflammation and so on.There is also related literature about hepatotoxicity after taking it.The key technique of traditional vegetable cultivation in Lijiang area is systematically summarized in this paper for reference in the future cultivation process. | Genlin YANG Qinghua LI Yungui GAO Zhigang YANG | 2021 | Medicinal Plant2021，12，2： | 0 |
| 7 | Effect of ciliary neurotrophic factor on bcl-2 and c-jun gene and protein expression in cultured retinal nerve cells显示文摘BACKGROUND:In various retinal neurodegenerative animal models,ciliary neurotrophic factor (CNTF) exhibits prominent neuroprotective effects on retinal nerve cells.Bcl-2 is an anti-apoptotic protein.c-Jun is upregulated and phosphorylated in the activated c-Jun N-terminal kinase pathway,which subsequently mediates apoptosis.However,the effect of CNTF on Bcl-2 and c-Jun expression in retinal nerve cells remains unclear.OBJECTIVE:To determine the dynamic changes in retinal nerve cell apoptosis,as well as bcl-2 and c-jun gene and protein expression,following a single dose of CNTF in a short period of time.DESIGN,TIME AND SETTING:A single-blind,randomized,controlled,in vitro experiment was performed at the Central Laboratory of Beijing Tongren Hospital from May 2008 to April 2009.MATERIALS:Neonatal bovine retinal nerve cells (Chinese Holstein),recombinant human CNTF (PeproTech,Rocky Hill,NJ,USA),rabbit polyclonal anti-Bcl-2 and c-Jun antibodies (Abcam,Cambridge,UK),fluorescein isothiocyanate-conjugated annexin V/propidium iodide kit (BioVision,Mountain View,CA,USA),real time polymerase chain reaction instrument (ABI,Foster City,CA,USA),and flow cytometer (BD FACSCalibur,Franklin Lakes,NJ,USA).METHODS:Neonatal bovine retinal cells from passage 2 were cultured for 3 days and incubated with,or without,50 ng/mL CNTF (control).MAIN OUTCOME MEASURES:Cell apoptosis was detected via Annexin V-FITC/PI double-staining and flow cytometry.bcl-2 and c-jun mRNA and protein expression were detected by quantitative real time polymerase chain reaction and western blot analysis.RESULTS:The proportion of late-stage apoptotic cells was significantly decreased at 2,4,and 6 days after CNTF treatment compared with the control group (P < 0.01).CNTF did not alter bcl-2 mRNA expression at the three time points,but significantly increased Bcl-2 protein expression at 2 and 4 days (P < 0.01).c-jun mRNA expression was significantly decreased 4 days after CNTF treatment (P < 0.01).In addition,c-Jun protein expression was slightly increased at 4 days (P < 0.01),but decreased at 6 days,compared with the control group (P < 0.05).CONCLUSION:A single dose of CNTF (50 ng/mL) upregulated Bcl-2 protein and downregulated c-jun mRNA expression,followed by a parallel,but lagged,change in c-Jun protein production in cultured neonatal bovine retinal nerve cells.These results suggested that CNTF reduces retinal nerve cell apoptosis by modifying Bcl-2 and c-Jun expression. | Xiang Zhang Xiang Lei Yueyue Liu Genlin Li | 2010 | Neural Regeneration Research2010，5，9： | 0 |
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