人类新基因HBVDNAPTP1结合蛋白的生物信息学分析     

伦永志  刘为国  朱莹  张国元  毛小强  司红敏  王琳 《大连大学学报》2006,27(6):80-83

[目的]应用生物信息学分析人类新基因乙型肝炎病毒(HBV)DNA聚合酶(Polym erase)反式调节蛋白(HBVDNAPTP1)结合蛋白(HBVDNAPTP1BP).[方法]利用生物信息学技术分析HBVDNAPTP1BP基因的染色体定位与组织表达,以及编码蛋白的化学物理性质与结构特征.[结果]HBVDNAPTP1BP基因染色体定位于1号染色体短臂3区5带1亚带,可在组织中低表达,但在多个组织中无表达.HBVD-NAPTP1BP的相对分子量为11 905.6,理论pI为7.72,不同条件下的消光系数为14 230 M-1cm-1或13 980M-1cm-1(280 nm),在体外哺乳动物网状细胞中的半寿期为30 h,并且无卷曲螺旋区域,但具有3个较强的疏水区域.HBVDNAPTP1BP无特殊二级结构,仅有1个蛋白激酶C磷酸化位点,不具有跨膜螺旋结构,无信号肽序列,定位于细胞核中.[结论]应用生物信息学对HBVDNAPTP1BP进行了分析,为进一步研究其生物学功能及其在乙型肝炎、肝细胞癌中的作用机制提供了依据和线索.  相似文献   

常量法和微量法测定蛋白质含量综合性实验设计     

赵玉荣  陈翠霞  张黎民  李幸凡  王文鑫 《实验技术与管理》2021,(2):44-48

蛋白质含量测定是生物化学和分子生物学研究中最常用的分析方法之一。该文以Bradford法为基础,利用紫外分光光度计(常量法)和多功能酶标仪(微量法)分别进行蛋白质含量测定。在此基础上,总结了常量法和微量法在测定蛋白质含量中的异同点。结果表明:常量法适合样品数量少、体积大的体系;而微量法则可以大大提高测试效率,特别适宜样品数量多、体积少的体系表征。由于微量法在相同条件下可同时完成多个样品的测试,具有更高的测量准确性。两种方法在实验中的综合运用可大大提高实验效率,有助于学生掌握Bradford法测定蛋白质含量的原理,认识常量法和微量法测定蛋白质含量的共性及特性,提高学生对紫外分光光度计和多功能酶标仪工作原理及工作特点的认识,显著提升学生的实验能力和逻辑思维能力,为从事相关科研工作奠定基础。  相似文献   

CHOP及GRP78在复发性多形性胶质母细胞瘤患者表达显著增加     

文雪 《孝感职业技术学院学报》2021,(1):108-112

目的:研究CHOP及GRP78与复发性GBM的相关性。方法:采用Western blot检测CHOP及GRP78在初发及复发GBM组织中的表达;Real-time PCR检测GRP78在初发以及复发GBM组织中的表达。结果:CHOP及GRP78在初发GBM中表达上调,而在经过原发性肿瘤切除后接受放疗和替莫唑胺化疗的复发性GBM中增加尤为显著。结论:CHOP及GRP78在GBM发病机制中起重要作用,该分子有望成为GMB潜在的诊断及预后评估标志物。  相似文献   

Mass spectrometry-based protein-protein interaction techniques and their applications in studies of DNA damage repair     

Zhen CHEN  Junjie CHEN 《Journal of Zhejiang University. Science. B》2021,22(1):1-20

Proteins are major functional units that are tightly connected to form complex and dynamic networks.These networks enable cells and organisms to operate properly and respond efficiently to environmental cues.Over the past decades,many biochemical methods have been developed to search for protein-binding partners in order to understand how protein networks are constructed and connected.At the same time,rapid development in proteomics and mass spectrometry(MS)techniques makes it possible to identify interacting proteins and build comprehensive protein-protein interaction networks.The resulting interactomes and networks have proven informative in the investigation of biological functions,such as in the field of DNA damage repair.In recent years,a number of proteins involved in DNA damage response and DNA repair pathways have been uncovered with MS-based protein-protein interaction studies.As the technologies for enriching associated proteins and MS become more sophisticated,the studies of protein-protein interactions are entering a new era.In this review,we summarize the strategies and recent developments for exploring protein-protein interaction.In addition,we discuss the application of these tools in the investigation of protein-protein interaction networks involved in DNA damage response and DNA repair.  相似文献   

CRISPR/Cas: a Nobel Prize award-winning precise genome editing technology for gene therapy and crop improvement     

Li  Chao  Brant  Eleanor  Budak  Hikmet  Zhang  Baohong 《Journal of Zhejiang University. Science. B》2021,22(4):253-284

Since it was first recognized in bacteria and archaea as a mechanism for innate viral immunity in the early 2010 s,clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated protein(Cas) has rapidly been developed into a robust, multifunctional genome editing tool with many uses. Following the discovery of the initial CRISPR/Cas-based system, the technology has been advanced to facilitate a multitude of different functions. These include development as a base editor, prime editor, epigenetic editor, and CRISPR interference(CRISPRi) and CRISPR activator(CRISPRa) gene regulators. It can also be used for chromatin and RNA targeting and imaging. Its applications have proved revolutionary across numerous biological fields, especially in biomedical and agricultural improvement. As a diagnostic tool, CRISPR has been developed to aid the detection and screening of both human and plant diseases, and has even been applied during the current coronavirus disease 2019(COVID-19) pandemic. CRISPR/Cas is also being trialed as a new form of gene therapy for treating various human diseases, including cancers, and has aided drug development. In terms of agricultural breeding, precise targeting of biological pathways via CRISPR/Cas has been key to regulating molecular biosynthesis and allowing modification of proteins, starch, oil, and other functional components for crop improvement. Adding to this, CRISPR/Cas has been shown capable of significantly enhancing both plant tolerance to environmental stresses and overall crop yield via the targeting of various agronomically important gene regulators. Looking to the future, increasing the efficiency and precision of CRISPR/Cas delivery systems and limiting off-target activity are two major challenges for wider application of the technology. This review provides an in-depth overview of current CRISPR development, including the advantages and disadvantages of the technology,recent applications, and future considerations.  相似文献   

裸子植物的生化系统学(三)——从种子蛋白多肽和针叶过氧化物酶探讨红豆杉科的系统位置     

胡志昂  王洪新  刘长江  潜忠兴 《中国科学院研究生院学报》1986,24(4):260-263

 SDS聚丙烯酰胺凝胶电泳揭示红豆杉属和白豆杉的种子蛋白主要多肽的分子量为31、   22和20千道尔顿(K)。穗花杉的多肽谱与上述两属相差较大，没有22K多肽，代之以一个33K   主要多肽。榧树属和穗花杉有很多共同的多肽，但没有44K这个红豆杉科多数种类共有的中   等含量的多肽，并出现一个36K主要多肽。三尖杉属一些种的多肽谱十分接近红豆杉属，竹   柏的种子蛋白与上述分类群也有一定程度的近似。     红豆杉科各属之间针叶过氧化物酶谱差别很大，但三尖杉属一些种与红豆杉属却有些类   似。     两种蛋白质资料一致说明红豆杉科内的进化趋势是从红豆杉属、经白豆杉属和穗花杉属   至榧树属。红豆杉属和三尖杉属之间蛋白质的近似，说明红豆杉科和三尖杉科之间的关系，可 能通过红豆杉属相联系的,述资料也说明红豆杉科应置于松柏目之下。  相似文献   

分子伴侣、蛋白质聚集和大分子拥挤环境对蛋白质折叠的影响     

李剑  王志珍 《中国科学院研究生院学报》2002,19(2):215-218

研究了细胞内存在的分子伴侣、蛋白质聚集和大分子拥挤环境对蛋白质折叠的影响.首先,发现分子伴侣GroEL与底物蛋白的结合有"半位"和"全位"两种模式,它是由底物蛋白的分子形状、分子大小以及与GroEL的相互作用性质决定的.接着,发现两种不同的蛋白质一起复性时相互不干扰,提示细胞内蛋白质折叠可能不受其他蛋白聚集的影响;后又发现α-乳清蛋白的前熔球态不仅是分子伴侣也是蛋白质聚集体的作用对象.最后,研究大分子拥挤环境对蛋白质折叠热力学和动力学的影响,揭示了这种影响的复杂性和多样性.  相似文献   

分子伴侣、蛋白质聚集和大分子拥挤环境对蛋白质折叠的影响(英)     

李剑  王志珍 《中国科学院研究生院学报》2002,(2)

研究了细胞内存在的分子伴侣、蛋白质聚集和大分子拥挤环境对蛋白质折叠的影响.首先,发现分子伴侣GroEL与底物蛋白的结合有 半位 "和 全位"两种模式,它是由底物蛋白的分子形状、分子大小以及与GroEL的相互作用性质决定的.接着,发现两种不同的蛋白质一起复性时相互不干扰,提示细胞内蛋白质折叠可能不受其他蛋白聚集的影响;后又发现α 乳清蛋白的前熔球态不仅是分子伴侣也是蛋白质聚集体的作用对象.最后,研究大分子拥挤环境对蛋白质折叠热力学和动力学的影响,揭示了这种影响的复杂性和多样性.  相似文献   

乳腺癌耐受蛋白介导5-氟脲嘧啶的耐受及机制探讨     

袁建辉  贺智敏  吕辉  余艳辉  陈主初 《深圳特区科技》2005,(Z1):106-112

AIM To filtrate breast cancer resistance protein(BCRP)-mediated resistance agents and investigate the mechanism,so as to provide valuable datum for optimization clinical chemotherapy scheme to tumor with evaluation marker of BCRP expression. METHODS MTT assay was used to filtrate BCRP-mediated resistance agents with PA317/Tet-on/TRE-BCRP cell of different expression levels of BCRP after treated with different concentration anticancer agents. High performance liquid chromatography(HPLC) was applied to measure relative dose of intracellular retention resistance agents. Nuclear DNA fluorescence dye,Hochest 33258, staining and flow cytometry were adopted to detect apoptotic cells after treated with drugs. RESULTS There were shown increasing durg-resistance to 5-fluorouracil,methotrexate, doxirubicin, pirarubicin,etoposide and mitoxantrone followed with increasing expression of BCRP on PA317/Tet-on/TRE-BCRP cells(P＜0.05, n=3),but shown sensitive to paclitaxel, cisplatin, vincristine, mitomycin and vindesine. There also was shown significant negative correlation between the intracellular retention dose of 5-fluorouracil with different expression of BCRP(r=-0.885, P＜0.05, n=3).There were shown parallel results of that decreasing cellular apoptotic rate with increasing cellular expression of BCRP after treated with 5-fluorouracil by fluorescence dye staining and flow cytometry(P＜0.05, n=3),and also shown significate rise of the apoptotic rate of BCRP expression cells after treated with Ko143 (P＜0.05, n=3). Every group of cells could be different extently blocked in phase of G0/G1 treated with 5-fluorouracil. CONCLUSION Resistance of 5-fluorouracil could be especially mediated by conjugated with BCRP and acted as drug exclude-pump substrate. Cellular ability resistant to 5-fluorouracil-induced apoptosis could be reinforced by BCRP expression.  相似文献   

玉兰减数分裂观察及染色体构型分析     

李建强  何子灿 《中国科学院研究生院学报》2003,41(4):362-368

采用去壁低渗方法，观察研究了玉兰Magnolia denudata有丝分裂和减数分裂的细胞学特征。实验结果证实玉兰存在两种染色体倍性，即2n=4x=76和2n=6x=114。通常，在木兰属甚至整个木兰科每个物种只具有一种染色体数目。玉兰有丝分裂间期核为复杂染色中心型，其中期染色体较小。玉兰在减数分裂中期I的构型表现出多样性，其中最主要的特点是比同源多倍体预期的二价体出现的频率更高些，其次是在减数分裂中期I可以观察到1或2个环状和（或）链状六价体。这些特征与同源异源六倍体或部分的异源六倍体种北美红杉Sequ  相似文献