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Advances in Microbiology 2021年第9期11卷 453-468页

作者： Mouna Ghorbel Ikram Zribi Najla Haddaji Malek Besbes Nouha Bouali Faiçal Brini Biology Department Faculty of Science University of Ha’il Ha’il Saudi Arabia Biotechnology and Plant Improvement Laboratory Centre of Biotechnology of Sfax (CBS) University of Sfax Sfax Tunisia

The pathogenesis-related proteins 1 (PR-1) gene family play important roles in the plant metabolism in response to biotic and abiotic stresses. The wheat TdPR1.2 has been previously isolated and characterized. Here we showed by bio-informatic analysis that TdPR1.2 contains six cysteine residues that are conserved between all PR-1 proteins tested. Using ScanProsite tool, we found that TdPR1.2 structure has a CRISP family signature 1 and 2 located at the C-terminal part of the protein. Those two domains are conserved in many identified PR1.2 proteins in plants. Moreover, SignalIP-5.0 analysis revealed that TdPR1.2 contains a putative signal peptide formed by 25 amino acids at the N-terminal extremity. The presence of this signal peptide suggested that the mature proteins will be secreted after the cleavage of the signal sequence. Further, we investigate the role of the TdPR1.2 proteins in the growth of Escherichia coli transformants cells under different abiotic stresses. Our results showed that the full-length form of TdPR1.2 enhanced tolerance of E. coli against salt and osmotic stress but not to KCl. Moreover, TdPR1.2 protein confers bacterial tolerance to heavy metals in solid and liquid mediums. Based on these results, we suggest that the TdPR1.2 protein could play an important role in response to abiotic stress conditions.

关键词： Abiotic Stress Bioinformatic Analysis Durum Wheat E. coli Growth Inhibition Pathogen Related proteins protein expression

Improved Catalytic Reaction of Biotemplated Palladium Nanoparticles through Immobilized Metal Affinity Purification

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Advances in Nanoparticles 2023年第1期12卷 22-31页

作者： Shadrach Ibinola Hazim Aljewari Imann Mosleh Robert Beitle Ralph E. Martin Department of Chemical Engineering University of Arkansas Fayetteville USA

To investigate the effect purification plays on nanoparticle (NP) synthesis and catalytic activity, three copies of Pd4 (TSNAVHPTLRHL) fused to the N-terminus of Green Fluorescent protein (GFP) was produced recombinantly and its characteristics pre and post purification was assessed. An E. coli expression system was employed, and purification was performed with Immobilized Metal Affinity Column (IMAC). Transmission electron microscopy (TEM) was utilized to examine the morphology of NPs synthesized with an enriched protein sample and ImageJ was used to determine the average size to be 2.44 nm. The turnover frequency of fabricated NP from the purified protein was analyzed by a model Suzuki-Miyaura coupling reactions and determined to be 33,000 hr-1. This value is three times higher than the turnover frequency when crude lysate containing (Pd4)3-GFP was used during NP synthesis. This result shows that enrichment enhanced the catalytic activity of NP.

关键词： protein expression Nanoparticle Synthesis Turnover Frequency Suzuki-Miyaura

Research on the Construction and Function of a Chlorophyll Degradation Recombinant Engineering Strain

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Agricultural Science & Technology 2017年第10期18卷 1788-1794页

作者： Zongcan YANG Zhan ZHANG Jianbin YE Yufu PENG Xiangzhen LIU Hongxia WANG Technology Center China Tobacco Henan Industrial Co. Ltd. School of Food and Biological Engineering Zhengzhou University of Light Industry

In order to effectively reduce the chlorophyll content in flue-cured tobacco, improve the overall quality of tobacco leaves, chlorophyllase gene was cloned from Arabidopsis thaliana. After the expression of the expression vector in E. coil, the recombinant engineering strain was obtained. Afterwards, IPTG （isopropy-β-D-thiogalactopyranoside）was used to induce the goal protein, and the chlorophyllase activity of the recombinant engineering strain was measured, so as to investigate its degradation effect on the chlorophyll in the extracts of tobacco leaves. The results were as follows：（1） the amplified chlorophyllase gene At- CLH1 constructed the expression vector pET28a-AtCLH1 successfully, obtaining the recombinant engineering strain; （2） induced under 30 ℃ for 22 h, the strain could well express the recombinant protein AtCLH1 with 0.5 mmol/L IPTG, and the molecular weight was about 35 kDa; （3） the strain showed good chlorophyllase producing capability, and the activity of the produced chlorophyllase could reach up to 24.9 U/mL, which could degrade the chlorophyll in tobacco extract and had a good application prospect in improving the quality of low quality tobacco; （4） based on the results of orthogonal test, the enzyme extract from the strain was added to the tobacco leaf surface, which could make the degradation rate of chlorophyll in the tobacco leaf reach 17.06% under the temperature of 37 ℃ at the humidity of 75% for 48 h; （5） after treated by the enzyme liquid, the test tobacco showed increase in the content of aromatic substances, enhancement of tobacco fragrance quality and amount, significant decrease of offensive odor and irritation, significant improvement of agreeable aftertaste, making the overall sensory quality of the tobacco leaf significantly improved.

关键词： Tobacco leaf Degradation of chlorophyll Recombinant Engineering strain protein expression Activity of chlorophyllase Orthogonal test Aroma substances Sensory quality

Comparison of RF3 B. junceato RF3 B. napus

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Food and Nutrition Sciences 2021年第11期12卷 1041-1061页

作者： Sandra J. W. Mackie BASF Corporation Research Triangle Park NC USA

RF3 Brassica napus L. (InVigorTM Canola) has been in commercial cultivation since 1997. RF3 B. napus was produced by Agrobacterium-mediated transformation to provide glufosniate ammonium herbicide tolerance to the crop. Canola quality (CQ) varieties of Brassica juncea L. (Czern & Coss), a close relative of B. napus, have been available in Canada for more than 20 years. Currently, BASF Agricultural Solutions is developing CQ RF3 B. juncea via conventional breeding with RF3 B. napus to provide similar herbicide tolerance in the B. juncea species. The insertion event RF3 contains the bar gene (origin Streptomyces hygroscopicus) coding for phosphinothricin acetyl transferase (PAT/bar) protein which confers tolerance to glufosinate-ammonium herbicides. RF3 also contains the barstar gene (origin Bacillus amyloliquefaciens), coding for the Barstar protein, which is an inhibitor of the Barnase protein. In the absence of Barnase, there is no impact of Barstar. Many safety studies have been conducted during the development of the canola quality lines of RF3 B. napus and RF3 B. juncea. This report seeks to compare the two species based on the scientific data generated during the safety assessments of RF3 B. napus and RF3 B. juncea and discusses the utility of using studies with one species to support the safety of the other species both containing the same insertion event.

关键词： Canola brassica juncea napus RF3 Composition protein expression Molecular Characterization

Insight of a novel functional gene related to differentiation of neural stem cells

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Journal of Shanghai University(English Edition) 2006年第5期10卷 465-468页

作者：庞淑亚赵翠萍文铁桥 Laboratory of Neural Molecular Biology School of Life Sciences Shanghai University Shanghai 200444 P.R. China

A novel fimctional gene P12 was isolated from neural stem cells cultured in differentiation medium. The fully length cDNA of P12 gene was cloned and sequenced. Result showed that it contains an open reading frame encoding a protein of 291 amino acids. Further, this gene was transferred into neural stem cell. Functional analysis suggests that the expression of p12 protein is closely correlated with differentiation of neural dendrite configuration. In addition, to obtain encoding protein, P12 sequence was also expressed in Pichia pastoris yeast.

关键词： neural stem ceils differentiation protein expression Pichia pastorfs.

Increased phosphorylation of CaMK Ⅱ in cerebral tissues of hypoxic preconditioned mice

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Increased phosphorylation of CaMK Ⅱ in cerebral tissues of ...

Identification of nPKCε-interacted proteins in the cortex of hypoxic preconditioned mice

中国病理生理学会受体、肿瘤和免疫专业委员会联合学术会议

作者：李海涛首都医科大学燕京医学院生理与病理生理研究室

Objective To explore the effect of hypoxic preconditioning(HPC) on middle cerebral artery occlusion(MCAO)-induced brain injury of mice and the changes of Calcium/calmodulin-dependent protein kinaseⅡ(CaMKⅡ) phosphoryl -ation and protein expression levels in the ischemic cortex. Methods Using our established HPC and MCAO mouse models,healthy male BALB/c mice weighing at 18～20 g were randomly divided into 4 groups:H0 sham,H4 sham,H0 and H4 *** blot combined with GelDoc imagine systems were applied to examine the changes in CaMKⅡphosphorylation and protein expression levels in the control,in the penumbra and the ischemic core of the brain from diffirent group *** Both phosphorylation and protein expression levels in the ischemic core and penumbra decreased significantly when compared with H0 sham group(P<0.05,n=6 for each group). But HPC increased the phosphorylation levels of CaMKⅡin the penumbra(P<0.05,n=6).Conclusion CaMKⅡmight involved in the attenuation of MCAO-induced brain injury by HPC of mice.

关键词： Cerebral hypoxic preconditioning MCAO CaMKⅡ Phosphorylation protein expression

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Identification of nPKCε-interacted proteins in the cortex o...

Differentially Expressed Proteome of Microcystis under Short-time Ultrasonic Stress

中国生理学会第23届全国会员代表大会暨生理学学术大会

作者： Su-Juan FENG,Nan ZHANG,Xuan YANG,Yan-Lin LUO,Song HAN,Jun-Fa LI~* Department of Neurobiology and Beijing Institute for Neuroscience,Capital Medical University,Beijing 100069,China

＜正＞Objective:Ischemic/hypoxic preconditioning（I/HPC） is an endogenous strategy in which sublethal hypoxic exposure induces the tissues/organs resistant to subsequent severe ischemic/hypoxic *** profound protective mechanism induced by I/HPC makes it an attractive target for potential clinical therapeutic approach, but the molecular mechanism underlying I/HPC is *** previous studies have demonstrated that the novel protein kinase Cε（nPKCε） participated in the development of cerebral ***,which proteins interacted with nPKCεfollowing HPC is *** this study,we try to identify nPKCε-interacted proteins in the cortex of HPC ***:The mouse HPC model was made according to our established *** decapitation, the separated cortex was rapidly homogenized in tissue lysis buffer to obtain cytosolic and particulate fractions. The techniques of immunoprecipitation（IP） and two-dimensional electrophoresis（2-DE） combining with ImageMaster 2D Platinum software were applied to analyze the differential expressions of nPKCε-interacted proteins;the target protein spots were identified by using matrix-associated laser desorption/ionization time of flight mass spectrometry（MALDI-TOF-MS） and Western ***:Compared with control group,there were 34 upregulated protein spots and 20 downregulated protein spots in cytosolic fraction,while 27 upregulated prtein spots and 28 downregulated protein spots were determined in particulate fraction of cerebral cortex of HPC mice. These target protein spots were identified by MALDI-TOF-MS and Swiss-prot database,and further distributed into categories according to their molecular functions and involvement in biological processes by using online PANTHER（protein Analysis Through Evolutionary Relationships） classification *** results showed that nPKCεcan interact with a variety of proteins such as some glycometabolism related proteins,heat shock proteins and cytoskeletal *** Western blot,we

关键词： ischemic/hypoxic preconditioning(I/HPC) novel protein kinase Cε(nPKCε) proteomics protein expression

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Agricultural Science & Technology 2017年第8期18卷 1371-1373,1415页

作者：骆灵喜李彬辉林秋月王波北京大学深圳研究院广东深圳518000 深圳市深港产学研环保工程技术股份有限公司广东深圳518000

This study was conducted to explore the regulation mechanism for key protein expression. The Microcystis treated by short-time ultrasonic wave was select-ed to analyze the total protein based on 2-DE. The results showed that there were 71 up-regulated protein spots, 56 down-regulated protein spots, 54 new protein spots and 21 protein spots disappeared under short-time ultrasonic stress. Eight dif-ferential proteins were chosen for further MALDI-TOFTOF/MS analysis, and the re-sults showed that 2 unknown proteins and 6 functional proteins were detected. These proteins were relevant to some physiological processes, such as antioxidation and anti-inflammatory process, phosphate synthesis and electron transfer, which is beneficial to the metabolic balance and self-protection under short-time ultrasonic stress.

关键词： Microcystis Short-time ultrasonic wave protein expression Two-dimen-sional electrophoresis

Angelica sinensis prevents apoptosis in ischemic penumbra Following middle cerebral artery occlusion in the rat

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中国临床神经科学 2000年第Z1期8卷 54-页

作者： ZHANG Junjtan HUANG Jianying ZHANG Xiaoqiang Department of Neurology Second Affiliated Hospital of Hubei Medical University Wuhan 430071 Department of Neurology Second Affiliated Hospital of Hubei Medical University Wuhan 430071 Department of Neurology Second Affiliated Hospital of Hubei Medical University Wuhan 430071

Objective To investigate the effect of Angelica sinensis on apoptosis in ischemic penumbra following transient cerebral ischemia. Methods The middl cerebral artery(MCA) of Wistar rats was occluded for 2 hours with the used of an intralaminal filament and recirculation was instituted for 48 hours. The Angelica sinensis were administered intracavitary 12g/kg .Apoptosis was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling(TUNEL). Bcl-2 and Bax pretien expression was detected by immunohistochemistry method. Results In the Angelica sinensis treated group,the apoptotic cells were significantly decreased (P＜0.01) and Bcl-2 protein expression were significantly increased (P＜0.01) than those in the control group in ischemic penumbra Conclusion Angelica sinensis prevents apoptosis in penumbra following cerebral ischemia may due to the enhanced expression of Bcl-2.

关键词： middle cerebral artery Angelica sinensis ischemic penumbra transient cerebral ischemia protein expression Wistar rats

THE RESEARCH ON MECHANISMS OF AMYLOID- β protein INDUCING ALZHEIMER'S DISEASE IN RAT AND EFFECTS OF NIMODIPINE TREATMENT

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中国临床神经科学 2000年第Z1期8卷 81-页

作者： Chunfeng Liu Shiyao Bao Yongping Dai Department of Neurology 2nd Affiliated Hospital Suzhou Medical College Department of Neurology 2nd Affiliated Hospital Suzhou Medical College Department of Neurology 2nd Affiliated Hospital Suzhou Medical College

0bjective In order to clarify the mechanism of the neurotoxics of Aβ, we studied the effects on the rCBF, extra-cellular amino acid (EAA), apoptosis and Bcl-2 protein expression, and their relationship with learning and memory deficiency. Methods Aβ was injected into NBM in rats to establish the AD model, learning and memory abilities were observed by Y-maze. The rCBF was measured by hydrogen clearance method. The EAA was detected by microdialysis in vivo with HPLC. The apoptosis and Bc1-2 protein expression was examined by flow cytometry. The male SD rats were divided into three groups: the model group was injected Aβ (10 μ g) into NBM of the rat. The control group was injected NaC1 (0.9%) in the same way. The treatment group was made with intraperitoneal injections of Nimodipine for 2 weeks after A β injections. Results The administration of Aβ into rat′s NBM could lead to the loss of learning and memory abilities. It was found that in frontal cortex and hippocampus, the rCBF decrease, however, with no trend of progressive decline. On the other hand, the levels of EAA increased, especially glutamate. Furthermore, A β significantly induced neurons apoptosis of frontal cortex and hippocampi cell, and upregulated the expression of the bcl-2. The Nimodipine might entirely improve rCBF of AD rats nearly to normal level, lessen the release of EAA and augment restrain neurotransmission. It might reduce the apoptosis partly, but it couldn't improve the learning and memory disorders completely. Conclusion The results implied that the neurotoxic effect of Aβ exists multi-mechanism.

关键词： learning and memory protein expression memory disorders flow cytometry In order