Kisspeptins在孕早期血浆和胎盘组织中的表达及其意义

目的:研究kisspeptins在孕早期血浆和胎盘组织中的表达,拟探讨其来源及其在妊娠中意义.方法:采用酶联免疫吸附法(ELISA),检测40例孕早期妇女和35例正常未妊娠妇女血浆kisspeptins的表达水平,且分析其与hCG的相关性;采用免疫组织化学方法进行其组织学定位.结果:(1)kisspeptins在妊娠组...     

Gene Expression Profile Changes in Germinating Rice

Water absorption is a prerequisite for seed germination. During imbibition, water influx causes the resumption of many physiological and metabolic processes in growing seed. In order to obtain more complete knowledge about the mechanism of seed germination, two‐dimensional gel electrophoresis was applied to investigate the protein profile changes of rice seed during the first 48 h of imbibition. Thirty‐nine differentially expressed proteins were identified, including 19 down‐regulated and 20 up‐regulated proteins. Storage proteins and some seed development‐ and desiccation‐associated proteins were down regulated. The changed patterns of these proteins indicated extensive mobilization of seed reserves. By contrast, catabolism‐associated proteins were up regulated upon imbibition. Semi‐quantitative real time polymerase chain reaction analysis showed that most of the genes encoding the down‐ or up‐regulated proteins were also down or up regulated at mRNA level. The expression of these genes was largely consistent at mRNA and protein levels. In providing additional information concerning gene regulation in early plant life, this study will facilitate understanding of the molecular mechanisms of seed germination.     

基因表达谱芯片的数据挖掘

随着基因芯片技术的迅速发展,表达谱芯片分析及aCGH等方法已被广泛应用于生命科学各个研究领域,由此产生的数据也呈指数级增长。如何从海量数据中获取有生物学意义的结果成为摆在生物学工作者面前的难题。对表达谱芯片数据挖掘方法进行了综述。介绍了基本分析思路,当前重点分析方向,如GO分析、pathway与调控网络分析、聚类分析等计算法则和相关几款易用的分析软件。并介绍了几种科学自由计算软件在表达谱生物信息学分析中的应用。藉此为从事芯片分析的研究人员提供参考。     

基因芯片表达谱数据的预处理分析

基因芯片数据的预处理是一个十分关键的步骤,通过数据过滤获取需要的数据、数据转换满足正态分布的分析要求、缺失值的估计弥补不完整的数据、数据归一化纠正系统误差等处理为后续分析工作做准备,预处理分析的重要性并不亚于基因芯片的后续分析,它将直接影响后续分析是否能得到预期的结果.本文重点综述了cDNA芯片的数据预处理,简要地概述寡核苷酸芯片的数据预处理.     

c—Ha-ras在转基因小鼠模型C57-ras各组织中的表达谱分析

目的：分析转基因c-Ha-ras在C57-ras癌症小鼠模型中的组织表达谱及时空表达差异。方法：利用半定量及荧光定量RT-PCR法,分析不同首建鼠系、不同周龄小鼠各脏器中转基因c-Ha-ras的表达。结果：转基因c-Ha-ras在心、肝等13种组织器官中均有表达,在肺脏中表达最高,在肝脏中表达最低,No.2、No.3和No.5等3个首建鼠均呈现相似的变化规律;转基因在No.5首建鼠中表达水平最高,而在同一个首建鼠系中,12周龄时表达高于8周龄和24周龄。结论：转基因c-Ha-ras在各脏器中能高效表达,并间接表明该转基因能稳定遗传,为C57-ras癌症小鼠模型用于新药临床前致癌性评价提了供理论支持。     

水稻低温响应基因OsCR1的克隆与表达分析

低温、高温、干旱等非生物胁迫是影响水稻产量与品质的重要非生物逆境因子.为了探索水稻耐逆的分子机理并挖掘新的水稻耐逆基因,采用Affymetrix 60K水稻基因表达芯片分析了培矮64S全基因组在上述逆境下的表达谱变化,筛选出一个受低温诱导表达水平显著升高的基因OsCR1( Oryza sativaL.cold resp...     

Gene Expression Profile Following Stable Expression of the Cellular Prion Protein

1. The cellular prion protein (PrPC) is expressed widely in neural and nonneural tissues at the highest level in neurons in the central nervous system (CNS).2. Recent studies indicated that transgenic mice with the cytoplasmic accumulation of PrPC exhibited extensive neurodegeneration in the cerebellum, although the underlying mechanism remains unknown. To identify the genes whose expression is controlled by overexpression of PrPC in human cells, we have established a stable PrPC-expressing HEK293 cell line designated P1 by the site-specific recombination technique.3. Microarray analysis identified 33 genes expressed differentially between P1 and the parent PrPC-non-expressing cell line among 12,814 genes examined. They included 18 genes involved in neuronal and glial functions, 5 related to production of extracellular matrix proteins, and 2 located in the complement cascade.4. Northern blot analysis verified marked upregulation in P1 of the brain-specific protein phosphatase 2A beta subunit (PPP2R2B), a causative gene of spinocerebellar ataxia 12, and the cerebellar degeneration-related autoantigen (CDR34) gene associated with development of paraneoplastic cerebellar degeneration.5. These results indicate that accumulation of PrPC in the cell caused aberrant regulation of a battery of the genes important for specific neuronal function. This represents a possible mechanism underlying PrPC-mediated selective neurodegeneration.     

Expression Profile of the REG Gene Family in Colorectal Carcinoma

Regenerating (REG) gene family belongs to the calcium-dependent lectin gene superfamily and encodes small multifunctional secretory proteins, which might be involved in cell proliferation, differentiation, and carcinogenesis. To clarify REG expression profile in colorectal carcinoma (CRC), the authors examined the expression of REG Iα, Iβ, III, HIP/PAP, and REG IV by immunohistochemistry on tissue microarray. The expression of REG Iα, III, and HIP/PAP was more frequently observed in the CRCs than adjacent non-neoplastic mucosa (p < 0.001), whereas it was the converse for REG Iβ and IV (p < 0.001). The expression of REG Iα, Iβ, III, and HIP/PAP was negatively correlated with the depth of invasion of CRCs (p < 0.05). The REG Iβ and HIP/PAP were less expressed in CRCs with than without venous invasion (p < 0.05). The positive rates of REG Iα and HIP/PAP were significantly higher in CRCs without than with lymph node metastasis (p < 0.05). Mucinous carcinoma more frequently expressed REG IV protein than well- and moderately differentiated ones (p < 0.05). There was a positive relationship between REG Iα, Iβ, III, and HIP/PAP expression (p < 0.05). Survival analysis indicated the REG Iβ or HIP/PAP expression was positively linked to favorable prognosis of carcinoma patients (p < 0.05). This study indicated that aberrant REG expression might be closely linked to the pathogenesis, invasion, or lymph node metastasis of CRCs. REG Iβ and HIP/PAP could be considered reliable markers of favorable prognosis of CRC patients.     

基因表达谱芯片在肿瘤基因组学中的应用

表达谱芯片 ,是指将几千个基因特异的探针或其ｃＤＮＡ片段固定在一块基因芯片上 ,对来源于不同个体 (正常人与患者 )、不同组织、不同细胞周期、不同发育阶段、不同病变、不同刺激 (包括不同诱导、不同治疗手段 )下的细胞内的ｍＲＮＡ或逆转录产物ｃＤＮＡ进行检测从而大规模对这些基因表达的个体特异性、组织特异性、发育特异性、分化阶段特异性、病变特异性、刺激特异性进行综合的分析和判断。基因表达谱芯片研究基因在不同组织或细胞、不同发育阶段中基因表达的改变 ,通过基因表达的改变进而阐明基因的功能。1 .杂交信号检测原理用不同…     

面向药物发现和精准医疗的基因表达谱分析

作为功能基因组学中重要的组成部分,基因表达谱在生物学、医学和药物研发等多个领域发挥着重要作用.特别是随着精准医疗概念的提出,整合多组学数据用于个性化医疗是未来的发展趋势.本文从基因表达谱的基本概念出发,重点介绍面向药物发现的基因表达谱分析方法,即基于关联图谱的方法、基于基因调控网络的方法和基于多组学数据整合的方法.系统整理了各种方法的研究进展,特别是在抗癌药物研发领域的最新进展,为利用基因表达谱数据进行药物研发提供方法借鉴.     

人类ZNF434基因的克隆、真核表达和组织表达谱分析

目的:克隆人类ZNF434基因并构建其真核表达质粒,鉴定ZNF434基因的组织表达谱。方法:实时定量PCR检测ZNF434基因在人体10种组织中的表达情况;克隆ZNF434基因的全长开放读码框区并连入真核表达载体pIRES2-EGFP,磷酸钙沉淀法转染HEK293T细胞,荧光显微镜和Western blot鉴定ZNF434蛋白的表达情况。结果:ZNF434基因在检测的人体组织中均有表达,其中骨髓和睾丸表达最高,成功克隆了ZNF434基因并构建了该基因的真核表达质粒ZNF434-pIRES2-EGFP,转染HEK293T细胞可观察到绿色荧光蛋白表达,Western blot可检测出分子量56kDa的Flag-ZNF434融合蛋白表达。结论:明确了人类ZNF434基因的组织表达谱,构建了该基因的真核表达载体,为该基因的进一步研究奠定了基础。     

应用cDNA微阵列研究肿瘤基因表达谱

随着研究水平的深入 ,对基因在发育过程中、生理反应中和疾病发展中表达上的时空差别 ,基因蛋白质产物的亚细胞定位和细胞分子间的相互作用等研究领域的开拓 ,所涉及到的基因的广泛性用传统方法研究已很难达到要求 ,因此一些新的研究方法应运而生 ,基因表达谱的研究即为其中之一。基因表达谱 ( ｇｅｎｅｅｘｐｒｅｓｓｉｏｎｐｒｏｆｉｌｅ)是指细胞中所有基因表达的格局 ,具体说来就是指细胞中产生所有ｍＲＮＡ的总和。对于所有的ｍＲＮＡ来说 ,通常采用的研究方法是将其逆转录成ｃＤＮＡ ,通过构建ｃＤＮＡ文库 ,再进行分析研究。原先采用…     

妊娠兔胎盘细胞凋亡及凋亡相关基因Bcl-2和Bax表达的动态变化

以新西兰雌兔为动物模型。研究妊娠期间胎盘细胞凋亡及其凋亡调控蛋白Bcl-2和Bax表达的动态变化,基因组DNA凝胶电泳实验检测到妊娠中期和晚期胎盘基因组DNA中出现典型的凋亡特征-DNA梯带,而且DNA断裂值在妊娠早、中、晚期分别为：0.14,0.49和1.43,与妊娠早期相比,妊娠中,晚期胎盘基因组DNA断裂值有显著性增加,TUNEL实验和活化caspase-3的免疫定位实验表明,在妊娠早期胎盘中存在细胞凋亡,而且在各妊娠期中细胞凋亡主要发生于合体滋养层,免疫印迹法分析表明,Bcl-2和Bax随妊娠的进行其表达量明显增加,Bax:Bcl-2比值在妊娠早、中、晚期分别为：0.89,0.91和1.25,呈增加趋势,实验结果说明,在兔正常妊娠中,胎盘合体滋养层细胞发生凋亡,且随妊娠的进行,凋亡细胞数量增多,胎盘细胞凋亡主要与细胞中Bax:Bcl-2的比例相关。