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Identification of the lysine and histidine transporter family in Camellia sinensis and the characterizations in nitrogen utilization 被引量:1
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作者 Wei Huang Danni Ma +9 位作者 Fawad Zaman Xulei Hao Li Xia E Zhang Pu Wang Mingle Wang Fei Guo Yu Wang Dejiang Ni Hua Zhao 《Horticultural Plant Journal》 SCIE CAS CSCD 2024年第1期273-287,共15页
In plants,the lysine and histidine transporter(LHT)family represent a class of proteins that mediate the uptake,translocation,and utilization of amino acids.The tea plant(Camellia sinensis)is a perennial evergreen wit... In plants,the lysine and histidine transporter(LHT)family represent a class of proteins that mediate the uptake,translocation,and utilization of amino acids.The tea plant(Camellia sinensis)is a perennial evergreen with a relatively high level of amino acids.However,systematic identification and molecular characterization of the LHT gene family has rarely been reported in tea plants.In this study,22 CsLHTs were identified from the‘Shuchazao’genome and classified into two groups.The modeled three-dimensional structure and the conserved domains presented a high similarity among the LHTs proteins.Moreover,it was predicted that a few genes were conserved through the analysis of the physiochemical characters,structures and cis-elements in promoters.The expression patterns in tea plants revealed that CsLHT7 was mainly expressed in the roots,and CsLHT4 and CsLHT11 exhibited relatively high expression in both the roots and leaves.Moreover,the expression of all three genes could be induced by organic nitrogen.Additionally,heterogeneous expression of CsLHT4,CsLHT7 and CsLHT11 in Arabidopsis thaliana decreased the aerial parts biomass compared with that in WT plants while significantly increased the rosette biomass only for CsLHT11transgenic plants versus WT plants.Overall,our results provide fundamental information about CsLHTs and potential genes in N utilization for further analysis in tea plants. 展开更多
关键词 Camellia sinensis Nitrogen lysine and histidine transporter(LHT)family
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Wetting alternating with partial drying during grain filling increases lysine biosynthesis in inferior rice grain
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作者 Yi Jiang Wenli Tao +2 位作者 Weiyang Zhang Zhiqin Wang Jianchang Yang 《The Crop Journal》 SCIE CSCD 2024年第1期262-270,共9页
Lysine content is a criterion of the nutritional quality of rice.Understanding the process of lysine biosynthesis in early-flowering superior grain(SG)and late-flowering inferior grain(IG)of rice would advance breedin... Lysine content is a criterion of the nutritional quality of rice.Understanding the process of lysine biosynthesis in early-flowering superior grain(SG)and late-flowering inferior grain(IG)of rice would advance breeding and cultivation to improve nutritional quality.However,little information is available on differences in lysine anabolism between SG and IG and the underlying mechanism,and whether and how irrigation regimes affect lysine anabolism in these grains.A japonica rice cultivar was grown in the field and two irrigation regimes,continuous flooding(CF)and wetting alternating with partial drying(WAPD),were imposed from heading to the mature stage.Lysine content and activities of key enzymes of lysine biosynthesis,and levels of brassinosteroids(BRs)were lower in the IG than in the SG at the early grainfilling stage but higher at middle and late grain-filling stages.WAPD increased activities of these key enzymes,BR levels,and contents of lysine and total amino acids in IG,but not SG relative to CF.Application of 2,4-epibrassinolide to rice panicles in CF during early grain filling reproduced the effects of WAPD,but neither treatment altered the activities of enzymes responsible for lysine catabolism in either SG or IG.WAPD and elevated BR levels during grain filling increased lysine biosynthesis in IG.Improvement in lysine biosynthesis in rice should focus on IG. 展开更多
关键词 BRASSINOSTEROIDS Inferior grain lysine biosynthesis Rice Wetting alternating with partial drying
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Effect of exogenous free N^(ε)-(carboxymethyl)lysine on diabetes-associated cognitive dysfunction:neuroinflammation,and metabolic disorders
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作者 Huang Zhang Lan Mo +8 位作者 Xi Chen Mei Li Maiquan Li Yang Xu Maomao Zeng Zhiyong He Qingwu Shen Jie Chen Wei Quan 《Food Science and Human Wellness》 SCIE CAS CSCD 2024年第5期2970-2981,共12页
Diabetes-associated cognitive dysfunction has already been attracted considerable attention.Advanced glycation end products(AGEs)from daily diets are thought to be a vital contributor to the development of this diseas... Diabetes-associated cognitive dysfunction has already been attracted considerable attention.Advanced glycation end products(AGEs)from daily diets are thought to be a vital contributor to the development of this diseases.However,the effect of one of the best-characterized exogenous AGEs N^(ε)-(carboxymethyl)lysine(CML)on cognitive function is not fully reported.In the present study,diabetical Goto-Kakizaki(GK)rats were treated with free CML for 8-weeks.It was found that oral consumption of exogenous CML significantly aggravated diabetes-associated cognitive dysfunction in behavioral test.In details,exogenous CML increased levels of oxidative stress,promoted the activation of glial cells in the brain,up-regulated the release of inflammatory cytokines interleukin-6,inhibited the protein expression of the brain-derived neurotrophic factor and thus led to neuroinflammation.Furthermore,exogenous CML promoted the amyloidogenesis in the brain of GK rats,and inhibited the expression of GLUT4.Additionally,several tricarboxylic acid cycle and glutamate-glutamine/γ-aminobutyric acid cycle intermediates including pyruvate,succinic acid,glutamine,glutamate were significantly changed in brain of GK rats treated with exogenous free CML.In conclusion,exogenous free CML is a potentially noxious compounds led to aggravated diabetes-associated cognitive dysfunction which could be possibly explained by its effects on neuroinflammation,energy and neurotransmitter amino acid homeostasis. 展开更多
关键词 Diabetes Cognitive dysfunction N^(ε)-(carboxymethyl)lysine NEUROINFLAMMATION METABOLOMIC
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Effects of Supplemental Glutamine and Lysine on Growth Performance of Broiler Chickens
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作者 Ali F. Alsogair Naif M. Alhawiti Samuel N. Nahashon 《Open Journal of Animal Sciences》 2024年第2期101-122,共22页
The optimum levels of Lysine and Glutamine needed for growth performance and maintenance of the chicken broilers were evaluated in a randomized 3 × 4 factorial arrangement of dietary treatments. The battery cages... The optimum levels of Lysine and Glutamine needed for growth performance and maintenance of the chicken broilers were evaluated in a randomized 3 × 4 factorial arrangement of dietary treatments. The battery cages measured 99 × 66 × 25 cm that can be sufficient for 5 birds. Day old Chicken broilers totaling 180 were assigned to dietary treatments comprising of 3 concentrations of Lysine (0.85, 1.14, and 1.42) each in combination with 4 concentrations of Glutamine (0, 1, 2, and 3). Each dietary treatment was replicated 3 times and each replication had 5 birds. The birds were given feed and water ad libitum with a 23-hour light regimen for a period of 4 weeks. Then, the experimental birds were evaluated for body weight gain, feed consumption, and feed conversion in order to determine their optimum requirement for dietary Lysine and Glutamine. Based on the findings of this study, the highest performance was observed in birds fed the diet supplemented with 1.42 lysine and 1% glutamine, but the highest improvement in feed conversion was observed in diet contain 1.14 and 1.42 with 1% and 3% glutamine, respectively. Birds fed 1.42 lysine and 1% glutamine had the highest total body weight gain and feed consumption. The lysine requirements in the diet for Chicken are between 1.14 and 1.42 with glutamine level of 1%. 展开更多
关键词 Broiler Chickens lysinE GLUTAMINE Amino Acid Dietary Protein Essentials AAs Non-Essential Amino Acids
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Feeding Practices and Use of Lysine and Methionine in Pigrationing on Intensified Pig Breeding in the West Center and Hauts Bassins Regions of Burkina Faso
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作者 Isidor Zangbéwindin Ouedraogo Timbilfou Kiendrebeogo +1 位作者 Aboubacar Toguyeni Chantal Yvette Kabore-Zoungrana 《Open Journal of Animal Sciences》 2024年第1期23-38,共16页
The aim of the study was to assess feeding practices and the use of lysine and methionine in pig rationing on intensified and semi-intensive pig breeding in the Koudougou and Bobo-Dioulasso areas. To this end, a cross... The aim of the study was to assess feeding practices and the use of lysine and methionine in pig rationing on intensified and semi-intensive pig breeding in the Koudougou and Bobo-Dioulasso areas. To this end, a cross-sectional survey was carried out on 87 breeding in these towns. A Discriminant Factorial Analysis (DFA) confirming a k-means classification of the data collected was used to retain 71 breeding divided into three breeding classes: Class A (32.4% of breeding), Class B (14.08%) and Class C (53.52%). The results show that the majority of pig breeders were men between the ages of 36 and 59. Average herd sizes were 35 ± 28;79 ± 42 and 89 ± 21 pigs for Classes A, B and C respectively. The main breeds of pig found on the breeding were crossbred, Large white, local, Landrace and Duroc. Class A (26.1%), B (30%) and C (15.8%) breeders were familiar with both lysine and methionine. Class A breeders distributed feed staggered (65.2%) and in rations (34.8%). Lysine (13%) and methionine (8.7%) were purchased at 5250 FCFA/kg. Those in class B distributed feed staggered (50%) and in the form of rations (50%), in which they incorporated lysine (30%) and methionine (30%) purchased at a cost of 2500 FCFA/kg and 3000 FCFA/kg respectively. Rationing and staggered feeding were practiced by 23.7% and 76.3% of Class C breeders respectively. Only lysine purchased at 3400 FCFA/kg was incorporated into rations by 10.5% of breeders. The high cost of lysine and methionine was incriminated by Class A (100%), B (33.3%) and C (50%) breeders. In conclusion, intensive pig breeding, the practice of rationing and the incorporation of the amino acids lysine and methionine are of ascending importance from classes C, A to B. The high cost of feedstuffs, particularly lysine and methionine, compromises their use in rations, which could have a negative impact on expected breeding performance. The screening and use of feeds rich in and/or enriched with these amino acids, through the development or adaptation of technologies, could improve the efficiency of rations and the productivity of intensive pig breeding in Burkina Faso. 展开更多
关键词 Feeding Practices lysinE METHIONINE Intensive Pig Breeding Burkina Faso
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Transfer of Lysine-rich Protein Gene into Rice and Production of Fertile Transgenic Plants 被引量:54
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作者 高越峰 荆玉祥 +3 位作者 沈世华 田世平 匡廷云 Samuel S.M.SUN 《Acta Botanica Sinica》 CSCD 2001年第5期506-511,共6页
Lysine-rich protein gene (lys) was cloned from Psophocarpus tetragonolobus (L.) DC. A plant expression plasmid was constructed and lys gene was under the control of maize ubiquitin promoter which is the highest effici... Lysine-rich protein gene (lys) was cloned from Psophocarpus tetragonolobus (L.) DC. A plant expression plasmid was constructed and lys gene was under the control of maize ubiquitin promoter which is the highest efficient monocotyledon promoter. The plasmid was introduced into rice embryogenic calli by microprojectile bombardment. The regenerated fertile plants were obtained by effective selection for hygromycin B resistance. Genomic PCR and Southern blotting analyses showed that the lys gene has been integrated into rice genome. Simultaneously, the results of GUS histochemical assay demonstrated that gus report gene is also expressed in leaves, stems and roots of the transgenic rice plants. Data analysis showed that lysine content in most of the 11 transgenic plants is differently improved, and in one of them increased by 16.04%. 展开更多
关键词 lysine-rich protein gene microprojectile bombardment transgenic rice lysine content
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中华乌塘鳢NK-lysin基因的克隆鉴定、表达分析及抗菌功能研究
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作者 徐婧 卢德政 +1 位作者 李昊霖 沈斌 《浙江海洋大学学报(自然科学版)》 CAS 2024年第4期277-287,共11页
NK-lysin是一种具有广谱抗菌活性和免疫调节功能的阳离子抗菌肽。通过分子克隆方法获得中华乌塘鳢的NK-lysin基因(BsNKL)cDNA序列。BsNKL编码152个氨基酸,包含1个信号肽、1个SapB结构域和6个保守半胱氨酸残基。基因表达分析表明,BsNKL... NK-lysin是一种具有广谱抗菌活性和免疫调节功能的阳离子抗菌肽。通过分子克隆方法获得中华乌塘鳢的NK-lysin基因(BsNKL)cDNA序列。BsNKL编码152个氨基酸,包含1个信号肽、1个SapB结构域和6个保守半胱氨酸残基。基因表达分析表明,BsNKL在主要器官组织中均有表达,副溶血弧菌感染能够引起BsNKL表达量在脾脏、头肾和鳃中发生显著变化。中华乌塘鳢BsNKL抗菌肽对金黄色葡萄球菌、大肠杆菌、副溶血弧菌和哈维氏弧菌等4种细菌均具有较好的抑菌活性。腹腔注射BsNKL能够为细菌感染的中华乌塘鳢提供有效的免疫保护并显著提高其存活率。综上所述,BsNKL在中华乌塘鳢抗细菌感染天然免疫应答方面发挥重要作用。 展开更多
关键词 中华乌塘鳢 抗菌肽 NK-lysin 基因表达 抑菌活性
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鸡抗菌肽NK-lysin的生物信息学分析 被引量:9
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作者 杭柏林 李杰 +3 位作者 张慧辉 徐彦召 徐军 胡建和 《黑龙江畜牧兽医》 CAS 北大核心 2017年第1期29-31,36,291,292,共6页
为了解鸡内源性抗菌肽NK-lysin的基本信息,试验选择鸡NK-lysin的氨基酸序列通过软件对NK-lysin的生物信息学进行了分析。结果表明:NK-lysin的等电点为5.87,分子质量为15 231.3 u,为稳定蛋白和亲水蛋白,定位于细胞质内,有1个跨膜区和1个... 为了解鸡内源性抗菌肽NK-lysin的基本信息,试验选择鸡NK-lysin的氨基酸序列通过软件对NK-lysin的生物信息学进行了分析。结果表明:NK-lysin的等电点为5.87,分子质量为15 231.3 u,为稳定蛋白和亲水蛋白,定位于细胞质内,有1个跨膜区和1个信号肽,没有糖基化位点,但有2个磷酸化位点,二级结构由α螺旋、延伸链、β折叠和无规则卷曲组成,存在多个抗原表位。说明鸡抗菌肽NK-lysin可作用于细胞膜或某些生物分子,使菌体死亡。 展开更多
关键词 抗菌肽 NK—lysin 生物信息学 二级结构 抗原表位
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厚壳贻贝M7lysin分子的克隆与表达 被引量:3
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作者 刘慧慧 薛超波 +1 位作者 常抗美 杨刚 《水产学报》 CAS CSCD 北大核心 2011年第9期1337-1342,共6页
M7 lysin位于贻贝精子顶体中,是溶解卵黄膜、促进受精作用的重要蛋白质,决定了贻贝种间精卵识别的特异性。采用同源克隆法得到厚壳贻贝M7 lysin分子,并在原核生物中对该分子进行重组表达。结果表明,扩增产物为540 bp左右的片段,进一步... M7 lysin位于贻贝精子顶体中,是溶解卵黄膜、促进受精作用的重要蛋白质,决定了贻贝种间精卵识别的特异性。采用同源克隆法得到厚壳贻贝M7 lysin分子,并在原核生物中对该分子进行重组表达。结果表明,扩增产物为540 bp左右的片段,进一步测序发现其开放阅读框cDNA为543 bp,与贻贝、地中海贻贝、盖勒贻贝具有较高的相似性;在线翻译所得蛋白质片段中含有180个氨基酸,分子量为20 ku,等电点为8.48;通过M7 lysin氨基酸序列构建系统进化树发现,贻贝和地中海贻贝亲缘关系最近,其次为盖勒贻贝,最后是厚壳贻贝;将M7lysin分子在E.coli Rosseta(DE3)中融合表达,得到包括载体氨基酸序列在内的25 ku蛋白质,符合预期分子质量。 展开更多
关键词 厚壳贻贝 精子 M7lysin 表达
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Expression of a Lysine-rich Gene in Bacillus subtilis 168 被引量:1
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作者 Li Xue-lin Liu Xiao-fei +3 位作者 Gao Xue-jun Qiao Bin Pan Hong-bao Ao Jin-xia 《Journal of Northeast Agricultural University(English Edition)》 CAS 2013年第1期43-48,共6页
Lysine-rich protein gene (lys) was cloned from winged bean (Psophocarpus tetragonolobus (L.) DC), and cloned into prokaryotie expression vector pHT43, the recombinant plasmid pHT43/lys were constructed and then ... Lysine-rich protein gene (lys) was cloned from winged bean (Psophocarpus tetragonolobus (L.) DC), and cloned into prokaryotie expression vector pHT43, the recombinant plasmid pHT43/lys were constructed and then transferred into Bacillus subtilis168, upon IPTG induction, the recombinant protein was expressed, and the content of lysine was detected by HPLC. The result showed that lysine content increased by 9.85%. It was suggested that introducing lys gene into Bacillus subtilis 168 was an effective way to improve its nutrition quality. 展开更多
关键词 lysine-rich protein gene Bacillus subtilis 168 lysine content HPLC
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Degradation kinetic study of lysine in lysine hydrochloride solutions for injection by determining its main degradation product 被引量:1
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作者 Mengying Tao Meng Zhu +1 位作者 Chunnuan Wu Zhonggui He 《Asian Journal of Pharmaceutical Sciences》 SCIE CAS 2015年第1期57-63,共7页
A limited number of researches have been reported to apply the Arrhenius equation to study the relationship between drugs and its degradation products so far.In the present work,the thermal degradation kinetics of lys... A limited number of researches have been reported to apply the Arrhenius equation to study the relationship between drugs and its degradation products so far.In the present work,the thermal degradation kinetics of lysine hydrochloride solutions for injection,the special solvent for ademetionine 1,4-butanedisulfonate(SAM)for injection,was investigated at selected temperatures and pH values.The main degradation product of lysine was separated,purified,and confirmed as lysine lactam.A reversed-phase high performance liquid chromatographic(RP-HPLC)method without derivation was developed for the simultaneous determination of lysine and lysine lactam.The results confirmed that both the lysine degradation and lysine lactam generation followed zero-order reaction kinetics.The degradation and generation rate constants increased with increasing temperatures and decreasing pH values.The temperature-dependent degradation and generation reaction could be sufficiently modeled on the Arrhenius equation with the activation energy of 80.14 and 83.22 kJ/mol,respectively.Meanwhile,a linear relationship existed between the amount of lysine degradation and lysine lactam generation since the approximate activation energy.Considering there could be other side effects,we established an upper limit of lysine lactam(500 mg/ml),as the acceptable criteria for stability to estimate the shelf life together with lysine,which made the prediction more accurate and credible.Extrapolation data demonstrated that the lysine hydrochloride solutions for injection could be stable for two years stored at room temperature. 展开更多
关键词 lysine hydrochloride lysine lactam Degradation kinetics Shelf life
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Rapid Determination of Lysine Content in Maize Kernel by Amino-acid Analyzer
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作者 齐建双 铁双贵 +3 位作者 韩小花 岳润清 燕树锋 卢彩霞 《Agricultural Science & Technology》 CAS 2015年第7期1340-1342,1397,共4页
In this study, nine high-lysine maize kernels and two kernels of common maize hybrid were used as experiment materials, and quantitative determination of lysine contents in high-lysine maize kernels and common maize k... In this study, nine high-lysine maize kernels and two kernels of common maize hybrid were used as experiment materials, and quantitative determination of lysine contents in high-lysine maize kernels and common maize kernels was carried out using Hitachi L-8900 Automatic Amino-acid Analyzer, to know the effect of the Analyzer in distinguishing the lysine contents between common maize kernels and high-lysine maize kernels. The results showed that the lysine contents of high-lysine maize kernels were among 0.34%-0.42%, while of common maize kernels were a- mong 0.24%-0.25%, and the difference was significant. Compared with other tradi- tional methods, this method is rapid, simple, sensitive, highly reproductive and needs fewer maize kernels, thus it is applicable in maize breeding. 展开更多
关键词 Amino-acid analyzer High lysine MAIZE
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团头鲂NK-lysin基因鉴定和表达分析 被引量:4
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作者 詹柒凤 丁祝进 +3 位作者 崔蕾 范君 王卫民 刘红 《水产学报》 CAS CSCD 北大核心 2016年第8期1145-1155,共11页
为研究抗菌肽NK-LYSIN在团头鲂免疫系统中的可能功能,本实验从团头鲂转录组中钓取到2个NK-lysin基因(nkla和nklb),通过PCR扩增并测序验证其ORF序列。通过荧光定量PCR检测其在健康组织和嗜水气单胞菌感染后免疫组织中的表达,并构建其原... 为研究抗菌肽NK-LYSIN在团头鲂免疫系统中的可能功能,本实验从团头鲂转录组中钓取到2个NK-lysin基因(nkla和nklb),通过PCR扩增并测序验证其ORF序列。通过荧光定量PCR检测其在健康组织和嗜水气单胞菌感染后免疫组织中的表达,并构建其原核表达体系。结果显示,团头鲂nkla和nklb分别编码122和136个氨基酸,NKLA和NKLB均属于鞘脂激活蛋白样蛋白家族成员,各含有6个高度保守的半胱氨酸及1个Sap B结构域。在健康团头鲂各组织中,nkla在脾脏中表达量极高,在肌肉和血液中表达量极低;而nklb则在肠中表达量较高,在头肾中表达量较低,在其他组织均有表达。nkla和nklb不同的表达模式暗示它们可能存在功能上的分化。经嗜水气单胞菌感染后,团头鲂nkla与nklb表达变化模式相似,头肾中,表达量在4 h时显著下降;肝脏中,表达量在4 h达到最高,而后回落到正常水平;在脾脏和肠中4 h时开始下降,72 h时达到峰值,暗示NK-lysin可能在团头鲂抗嗜水气单胞菌感染过程中发挥重要作用。另外,本实验对nkla和nklb进行了原核表达,为进一步研究NK-lysin的功能奠定了基础。 展开更多
关键词 团头鲂 嗜水气单胞菌 抗菌肽 NK-lysin 荧光定量 原核表达
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斑点叉尾NK-lysin成熟肽在毕赤酵母中的表达 被引量:5
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作者 吴慧 陶妍 《淡水渔业》 CSCD 北大核心 2017年第1期78-83,共6页
NK-lysin是毒性T淋巴细胞和自然杀伤细胞产生的具有抗菌作用的多肽,在机体免疫系统中发挥重要作用,被认为是抗生素的理想替代品。NK-lysin的生物学功能主要由其成熟肽(m NK-lysin)决定。本文在已有斑点叉尾(Ictalurus punctatus)NK-ly... NK-lysin是毒性T淋巴细胞和自然杀伤细胞产生的具有抗菌作用的多肽,在机体免疫系统中发挥重要作用,被认为是抗生素的理想替代品。NK-lysin的生物学功能主要由其成熟肽(m NK-lysin)决定。本文在已有斑点叉尾(Ictalurus punctatus)NK-lysin成熟肽原核重组表达载体p ET-32a(+)-m NK-lysin的基础上,通过PCR在m NK-lysin的5’端和3’端分别添加EcoRI和Xba I酶切位点,并在3’端添加6×His标签以便于纯化;扩增到的片段与表达载体p PICZαA连接构建重组表达载体p PICZαA-m NK-lysin,转化至毕赤酵母X-33细胞中,通过博来霉素筛选以及对酵母转化子基因组的PCR鉴定得到高拷贝酵母转化子;在29℃,250 rpm,采用0.5%甲醇进行诱导表达;表达产物经固化金属离子亲和层析(IMAC)获得纯化的重组体m NK-lysin。经Tricine-SDS-PAGE分析,其分子量约为11.5 k D;经Western blot分析,证明m NK-lysin在毕赤酵母中成功表达。 展开更多
关键词 斑点叉尾鮰(Ictalurus punctatus) NK-lysin成熟肽 毕赤酵母 重组DNA表达
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猪重组NK-lysin抑制肝细胞性肝癌的转移:基于下调FKBP3表达、抑制氧化磷酸化和糖酵解 被引量:1
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作者 范艺凡 冯志伟 +5 位作者 范阔海 尹伟 孙娜 孙盼盼 孙耀贵 李宏全 《南方医科大学学报》 CAS CSCD 北大核心 2023年第7期1116-1126,共11页
目的应用生物信息学方法探究猪重组NK-lysin(prNK-lysin)抑制肝癌细胞转移潜在的作用靶点和通路。方法将肝癌细胞设置为空白对照组,PBS处理组和prNK-lysin处理组,37℃作用6 h后,应用高效液相色谱串联质谱对肽段进行鉴定,按照Foldchange=... 目的应用生物信息学方法探究猪重组NK-lysin(prNK-lysin)抑制肝癌细胞转移潜在的作用靶点和通路。方法将肝癌细胞设置为空白对照组,PBS处理组和prNK-lysin处理组,37℃作用6 h后,应用高效液相色谱串联质谱对肽段进行鉴定,按照Foldchange=1.2倍且P<0.05筛选出差异表达蛋白。基于GO、KEGG等数据库对差异表达蛋白进行GO功能分析和KEGG通路分析。运用RT-qPCR验证细胞中多肽-N-乙酰半乳糖胺基转移酶13(GALNT13)、跨膜蛋白51(TMEM51)和FKBP脯酰异构酶3(FKBP3)的mRNA相对表达量。Western blot验证FKBP3的蛋白表达量。结果蛋白组学表明,与空白对照组相比,prNK-lysin处理组中有1989个差异表达蛋白;与PBS处理组相比,prNK-lysin处理组中有2753个差异表达蛋白;PBS处理组和空白对照组相比,有15个差异表达蛋白。相对于PBS处理组和空白对照组,prNK-lysin处理组中共有1909个差异表达蛋白。GO和KEGG分析表明差异表达蛋白主要参与Viral process、translational initiation、RNAbinding等过程,主要富集于Ribosome、Protein process in endoplasmic reticulum、RNA transport等通路。RT-qPCR表明,与空白对照组相比,prNK-lysin处理组显著升高了细胞内GALNT13(1.54±0.06 vs 1.02±0.17,P<0.05)和TMEM51(1.27±0.07 vs 1.00±0.04,P<0.01)的mRNA相对表达量,显著降低了FKBP3(0.43±0.06 vs 1.02±0.24,P<0.05)的mRNA相对表达量。Western blotting表明prNK-lysin处理组与空白对照组相比显著降低了细胞内FKBP3(0.68±0.02 vs 1.02±0.03,P<0.001)的蛋白表达量。结论prNK-lysin处理后肝癌细胞SMMOL/LC-7721的蛋白质组与空白组相比发生显著变化,prNK-lysin作用于FKBP3蛋白,并能通过影响细胞内氧化磷酸化和糖酵解等通路发挥其抑制作用。 展开更多
关键词 肝细胞性肝癌 蛋白组学 猪重组NK-lysin 差异表达蛋白
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斑点叉尾鮰NK-lysin基因的cDNA克隆及融合表达质粒构建 被引量:3
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作者 张卓娜 陶妍 《生物技术通报》 CAS CSCD 北大核心 2012年第9期166-172,共7页
以斑点叉尾鮰(Ictalurus punctatus)NK-lysin-type1的成熟肽为研究目标,首先通过RT-PCR和巢式PCR从斑点叉尾鮰的鳃中克隆到编码NK-lysin成熟肽的基因,该基因编码由92个氨基酸残基组成的成熟肽;6个高度保守的半胱氨酸残基位于成熟肽内,... 以斑点叉尾鮰(Ictalurus punctatus)NK-lysin-type1的成熟肽为研究目标,首先通过RT-PCR和巢式PCR从斑点叉尾鮰的鳃中克隆到编码NK-lysin成熟肽的基因,该基因编码由92个氨基酸残基组成的成熟肽;6个高度保守的半胱氨酸残基位于成熟肽内,它们被推测与NK-lysin的抗菌活性有关。为了进一步构建原核表达系统用于成熟肽的表达,pET-32a(+)被选择作为融合表达质粒,该质粒含有1个trxA融合头基因,与目的片段mNK-lysin连接后形成融合基因trxA-mNK-lysin,进而有助于在工程菌E.coliBL21(DE3)中的可溶性融合表达。通过PCR、EcoR I和HindⅢ双酶切处理以及DNA测序鉴定,证明pET-32a-mNK-lysin重组融合表达质粒已经被成功构建;将其转化至工程菌E.coliBL21(DE3)后的测序结果表明该重组质粒未发生任何DNA变异。 展开更多
关键词 斑点叉尾鮰 抗菌肽 NK-lysin CDNA克隆 融合表达质粒
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小球藻病毒PBCV-1特异性溶壁酶(Lysin)的溶壁活性 被引量:4
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作者 韩继刚 刘平芳 +2 位作者 康明 叶寅 田波 《微生物学报》 CAS CSCD 北大核心 2002年第3期354-358,共5页
从PBCV 1感染小球藻NC6 4A的细胞裂解液中提取了Lysin的粗制剂 ,酶活底物范围分析表明 ,几丁质酶、壳聚糖酶和 β 1 ,3 葡萄糖苷酶是Lysin活性的主要组成部分 ,并与小球藻细胞壁的组成成分相吻合。其中几丁酯酶和壳聚糖酶 ,特别是几丁... 从PBCV 1感染小球藻NC6 4A的细胞裂解液中提取了Lysin的粗制剂 ,酶活底物范围分析表明 ,几丁质酶、壳聚糖酶和 β 1 ,3 葡萄糖苷酶是Lysin活性的主要组成部分 ,并与小球藻细胞壁的组成成分相吻合。其中几丁酯酶和壳聚糖酶 ,特别是几丁酯酶在裂解小球藻细胞壁的过程中发挥了重要的作用。Lysin粗制剂经FPLC分离纯化得到分子量分别为 5 2kD、5 6kD的两个几丁质酶 (Chil和Chi2 )和一个分子量为 3 展开更多
关键词 几丁质酶 壳聚糖酶 β-1 3-葡萄糖苷酶 病毒侵染 小球藻病毒 PBCV-1 特异性溶壁酶 溶壁活性
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不同浓度脂多糖诱导绵羊免疫器官中NK-Lysin基因表达的研究 被引量:5
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作者 张殿卿 李蕊 +4 位作者 陈鹏博 赵庆亮 张国奇 李尤简 盛金良 《中国畜牧兽医》 CAS 北大核心 2014年第7期44-49,共6页
为研究病原体侵染动物机体时抗菌肽NK-Lysin在机体反应过程中所起到的防御作用及作用机理,本研究通过实时荧光定量PCR检测NK-Lysin在脂多糖(lipopolysaccharide,LPS)不同诱导条件下及不同免疫器官中的相对表达量,探讨绵羊抗菌肽NK-Lysi... 为研究病原体侵染动物机体时抗菌肽NK-Lysin在机体反应过程中所起到的防御作用及作用机理,本研究通过实时荧光定量PCR检测NK-Lysin在脂多糖(lipopolysaccharide,LPS)不同诱导条件下及不同免疫器官中的相对表达量,探讨绵羊抗菌肽NK-Lysin在免疫器官的生成和应答反应。构建目标基因NK-Lysin和内参基因GAPDH的克隆载体,绘制出标准曲线,检测NK-Lysin的表达量。结果显示,在LPS浓度为0.01、0.1、1、10μg/mL时,NK-Lysin mRNA表达量均在8h内达到最大值,NK-Lysin基因的表达量与LPS的浓度不存在正相关性;NK-Lysin mRNA在肩前淋巴结、脾脏及血液中均有不同程度的表达,脾脏中表达量与其他组织中的表达量相比差异极显著(P<0.01)。免疫器官能在较短时间内表达出抗菌肽NKLysin,使其参与机体先天性免疫应答反应。 展开更多
关键词 NK-1 ysin 实时荧光定量PCR 脂多糖 免疫器官 表达
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Effects of Dietary Net Energy Density and Standardized Ileal Digestible Lysine: Net Energy Ratio on the Performance and Carcass Characteristic of Growing-Finishing Pigs Fed Low Crude Protein Supplemented with Crystalline Amino Acids Diets 被引量:10
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作者 ZHANG Gui-jie YI Xue-wu +3 位作者 CHU Li-cui LU Ning Htoo John QIAO Shi-yan 《Agricultural Sciences in China》 CAS CSCD 2011年第4期602-610,共9页
Two 3×3 factorial design trials with 1 080 of Yorkshire×Landrace×Duroc barrows were conducted to investigate the effects of dietary net energy (NE) density and standardized ileal digestible (SID) ly... Two 3×3 factorial design trials with 1 080 of Yorkshire×Landrace×Duroc barrows were conducted to investigate the effects of dietary net energy (NE) density and standardized ileal digestible (SID) lysine:NE ratio on the performance of growing- finishing pigs and carcass characteristic of finishing pigs fed low crude protein (CP) supplemented with crystalline amino acids (CAA) diets. Low CP corn-soybean meal basal diets (14% for Exp. 1 and 11% for Exp. 2) supplemented with crystalline lysine, methionine, threonine, and tryptophan were formulated. Nine treatments with increasing NE density and SID lysine:NE ratio were used in each trial (Exp. 1: 9.66, 9.87, and 10.08 MJ kg-1 NE, 0.89, 0.96, and 1.03 g MJ-1 SID lysine: NE; Exp. 2: 9.83, 10.04, and 10.25 MJ kg-1 NE, 0.69, 0.76, and 0.83 g MJ-1 SID lysine:NE). In Exp. 1, we observed that the average daily gain (ADG) for barrows that received the diet containing 9.87 MJ kg-1 NE was significantly higher than that of barrows fed a 9.66 MJ kg-1 NE diet (P0.05), but no difference was observed when compared with barrows that received a diet containing 10.08 MJ kg-1 NE (P0.05). The ADG was affected both linearly (P0.05) and quadratically (P0.05) as the SID lysine:NE ratio increased. The NE density or SID lysine:NE ratio and their interaction had no effect on the average daily feed intake (ADFI) and serum urea nitrogen (SUN) (P0.05). The feed to gain ratio (F:G) decreased (linearly, P0.05; quadratically, P0.05) as the NE density or SID lysine:NE ratio increased, but no interaction of NE density and SID lysine: NE ratio on the F:G. In Exp. 2, the dietary NE density both linearly (P0.05) and quadratically (P0.05) affected the ADG and F:G. Barrows that received a diet containing 10.04 MJ kg-1 NE had a greater ADG (P0.05) than barrows that received a 9.83 MJ kg-1 diet, but there was no difference (P0.05) as compared with barrows that received a diet containing 10.25 MJ kg-1 NE. F:G was significantly higher (P0.05) for barrows that received a 9.83 MJ kg-1 diet when compared with barrows that received the other two diets. The ADG was significantly decreased (P0.05) for pigs that received a diet containing 0.69 g MJ-1 SID lysine:NE ratio when compared with pigs fed a diet containing 0.76 or 0.83 g MJ-1 SID lysine:NE ratio. The NE density or SID lysine:NE ratio and their interaction had no effect on the average daily feed intake, SUN, and carcass characteristic for finishing barrows (P0.05). The overall results of the two experiments indicated that barrows that received the diet with the dietary CP level reduced 4 percentage units compared the NRC (1998) recommendations. Maximized the best performance with NE density and SID lysine:NE ratio were 9.87 MJ kg-1, 1.03 g MJ-1 NE for 20-50 kg pigs, and 10.04 MJ kg-1, 0.76 g MJ-1 NE for 60-100 kg pigs, respectively. 展开更多
关键词 pig lysinE net energy growth performance carcass characteristic
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Study on Lysine and Methionine Content Promotion of Soybean Meal by Probiotic Fermentation Process 被引量:8
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作者 Lu Zhi-yong Gao Xue-jun +4 位作者 Huang Jian-guo Liu Rong Liu Ying Qiao Bin Qiu You-wen 《Journal of Northeast Agricultural University(English Edition)》 CAS 2012年第1期62-67,共6页
Soybean meal (SBM) is commonly used for livestock feeds, but its application in diets for livestock is limited due to some antinutritional factors. The contents of methionine and lysine of soybean meal were promoted... Soybean meal (SBM) is commonly used for livestock feeds, but its application in diets for livestock is limited due to some antinutritional factors. The contents of methionine and lysine of soybean meal were promoted by Bacillus natto and Leuconostoc mesenteroides fermentation, benefial for the livestock feeds. It was crude protein (CP) 56.8%, methionine 43.56 mg · g^-1, and lysine 74.87 mg · g^-1, cows fed a diet with FSBM milk yield raised 14.2%, the change in the milk protein, the lactose and the dry matter content had also obvious increase. This convenient technique offers helpful exploration for industrialization of soybean meal fermentation. 展开更多
关键词 soybean meal FERMENTATION Bacillus natto Leuconostoc mesenteroides METHIONINE lysinE
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