选用柱层析、电泳和反相高效液相色谱(RP-HPLC)技术制备质谱纯棕色固氮菌细菌铁蛋白(Bacteri-al ferritin ofAzotobacter vinelandii,AVBF),并采用释放铁动力学和肽质量指纹图谱(Peptide mass fingerprint-ing,PMF)技术分别鉴定A...
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选用柱层析、电泳和反相高效液相色谱(RP-HPLC)技术制备质谱纯棕色固氮菌细菌铁蛋白(Bacteri-al ferritin ofAzotobacter vinelandii,AVBF),并采用释放铁动力学和肽质量指纹图谱(Peptide mass fingerprint-ing,PMF)技术分别鉴定AVBF。基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)和电泳技术揭示AVBF亚基之间相互作用强度、稳定性和聚合态。AVBF可直接捕获有机小分子亚甲蓝(MB),其捕获率为15.0±2.0MB/AVBF,认为介于AVBF亚基单体之间的血红素参与捕获MB。较高浓度(40%;0%)的乙腈和丙酮均能使AVBF和鲨鱼肝铁蛋白(Liver ferritin of shark,SLF)释放不稳定亚基,但在较低浓度(20%;0%)的乙腈条件下,却需要借助来源于质谱仪的激光才能使AVBF或SLF释放不稳定亚基,并供质谱分析。AVBF亚基之间的相互作用强度明显低于SLF。铁蛋白亚基之间的相互作用强度高低与铁蛋白执行释放和储存铁的速率有关。
优化建立鲤鱼(cyprinus carpio,CC)嗅觉端脑(嗅脑)全蛋白提取技术。联用低渗透裂解和液氮冻溶法破碎鲤鱼嗅脑组织(rhinencephalon tissue of cyprinus carpio,CCRT)、低速离心提取CCRT全蛋白,并采用双向凝胶电泳(2D-PAGE)技术进行有效...
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优化建立鲤鱼(cyprinus carpio,CC)嗅觉端脑(嗅脑)全蛋白提取技术。联用低渗透裂解和液氮冻溶法破碎鲤鱼嗅脑组织(rhinencephalon tissue of cyprinus carpio,CCRT)、低速离心提取CCRT全蛋白,并采用双向凝胶电泳(2D-PAGE)技术进行有效分离。经分析与统计,每张CCRT的2D-PAGE图谱中的蛋白质斑点数目约为1200个。分别分离CCRT的脂溶性和水溶性全蛋白,并获得高分辨率的2D-PAGE图谱。选用差异蛋白质组学技术筛选经10%冰醋酸创伤后的CC,其端脑组织所表达出的6种应激蛋白质,并用肽质量指纹谱(peptide mass fingerprinting,PMF)和数据库检索技术给予鉴定。其中3种蛋白质为70S热休克蛋白、β微管蛋白和DNA链接酶IV,有望作为研究大脑急性创伤后的应激修复途径和机理的指示蛋白质。
To explore the existence and distribution of nucleophosmin in the nuclear matrix and its co-localization with the other related gene products following HMBA treatment in the human hepatocarcinoma SMMC-7721 cells,the n...
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To explore the existence and distribution of nucleophosmin in the nuclear matrix and its co-localization with the other related gene products following HMBA treatment in the human hepatocarcinoma SMMC-7721 cells,the nuclear matrix of SMMC-7721 cells was extracted pre/post HMBA induced differentiation.2D PAGE proteomics analyses showed that nucleophosmin existed in the fractions of nuclear matrix proteins and was down-regulated after HMBA treatment with further confirmation by Western blot *** immunofluorescence observation revealed that nucleophosmin located in the nuclear matrix,HMBA treatment altered its expression level and distribution *** co-localization of nucleophosmin with cancer-related genes and the products of oncogenes or tumor repression genes,including c-fos,c-myc,p53 and Rb,using laser scanning confocal microscopy,were evaluated,and substantial differences were observed following HMBA *** results implies that nucleophosmin,as a nuclear matrix protein,the level of its expression and the colocalization with cancer-related gene products may play an important role during the differentiation of SMMC-7721 cell.
为了研究铜绿微囊藻 Microcystis aeruginosa PCC7820中是否存在内源昼夜节律,检测了其细胞中的三磷酸腺苷(ATP)含量和细胞分裂的日变化规律。通过生物发光法检测的细胞内 ATP 含量变化结果表明,在12h光/12h暗(L/D)周期环境下该藻的 ATP...
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为了研究铜绿微囊藻 Microcystis aeruginosa PCC7820中是否存在内源昼夜节律,检测了其细胞中的三磷酸腺苷(ATP)含量和细胞分裂的日变化规律。通过生物发光法检测的细胞内 ATP 含量变化结果表明,在12h光/12h暗(L/D)周期环境下该藻的 ATP 含量呈现明显的近似24h 的昼夜周期性变化,且这种周期性变化在连续光照条件下能至少持续运行3个周期,周期长度具有温度补偿效应,而光照和温度的改变能重置昼夜节律的时相,这说明铜绿微囊藻 ATP 含量的昼夜节律变化受到内源生物钟的控制。通过对细胞数、细胞大小和细胞分裂素含量的检测发现,传代时间为38.4h 的铜绿微囊藻的细胞分裂也呈现受生物钟调控的昼夜节律性,这种昼夜节律变化可能是生物钟通过一种门控机制调控 *** PCC7820的细胞分裂时相而产生的。
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