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HBV基因定量分析技术的应用价值、存在问题及展望 被引量:30
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作者 缪晓辉 孔宪涛 《肝脏》 1999年第3期172-174,共3页
关键词 乙型肝炎 HBV 基因定量分析
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乙肝病毒基因定量分析的临床意义及应用价值 被引量:2
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作者 周小平 《中西医结合肝病杂志》 CAS 2003年第5期311-313,共3页
近年来乙肝病毒(HBV)基因定量分析的价值受到越来越多研究者的重视.以前由于过分强调了HBV感染过强的细胞免疫应答介导肝细胞损伤,因而忽视了对病毒复制水平与致病性之间因果关系的研究.
关键词 乙肝病毒 基因定量分析 临床意义 诊断 免疫应答 肝细胞损伤
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HBV基因定量分析技术的原理和方法 被引量:10
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作者 缪晓辉 戚中田 《肝脏》 1999年第1期46-48,共3页
关键词 HBV 基因定量分析 原理 方法 PCR 分子杂交
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颗粒酶/穿孔素定量分析与STR基因检测对移植早期GVHD诊断价值
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作者 邵媛 叶欣 +5 位作者 丁浩强 夏文杰 徐秀章 邓晶 王嘉励 陈扬凯 《广州医药》 2011年第6期48-50,共3页
目的比较颗粒酶/穿孔素基因表达定量分析和短串联重复序列(STR)基因位点检测对移植早期GHVD的诊断价值。方法 5例白血病患者在移植前和移植后1、3、5、7、9、11、14、30天采用荧光定量PCR检测颗粒酶/穿孔素和短串联重复序列检测的方法,... 目的比较颗粒酶/穿孔素基因表达定量分析和短串联重复序列(STR)基因位点检测对移植早期GHVD的诊断价值。方法 5例白血病患者在移植前和移植后1、3、5、7、9、11、14、30天采用荧光定量PCR检测颗粒酶/穿孔素和短串联重复序列检测的方法,并比较其检测结果。结果 5名患者颗粒酶/穿孔素基因水平不呈曲线变化,从术前到30天基因表达水平无明显差异。短串联重复序列基因位点检测从最初的供者基因嵌合状态到30天最后一次检测都是完全独立植入。结论短串联重复序列基因位点检测反应供者基因植入的情况,提示患者复发和GVHD的发生,对临床早期诊断和治疗有一定的指导意义。 展开更多
关键词 颗粒酶 穿孔素 基因定量分析 短串联重复序列(STR) 移植物抗宿主病(GVHD)
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人周围血单核细胞载脂蛋白E基因表达的竞争性逆转录-聚合酶链反应定量分析 被引量:1
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作者 向伟 马燕琳 +7 位作者 符生苗 赵水平 赵迪成 杨进福 陈炽 王福利 王果 聂赛 《中国动脉硬化杂志》 CAS CSCD 2003年第5期473-476,共4页
为建立人周围血单核细胞载脂蛋白E基因表达检测方法 ,研究载脂蛋白E基因与儿童健康的关系 ,我们抽取 2 6例健康儿童外周静脉血 ,分离血单核细胞 ,抽提RNA ,采用逆转录—聚合酶链式反应检测载脂蛋白E基因表达 ,并以正常人cDNA作定量标准... 为建立人周围血单核细胞载脂蛋白E基因表达检测方法 ,研究载脂蛋白E基因与儿童健康的关系 ,我们抽取 2 6例健康儿童外周静脉血 ,分离血单核细胞 ,抽提RNA ,采用逆转录—聚合酶链式反应检测载脂蛋白E基因表达 ,并以正常人cDNA作定量标准物 ,待测样品与定量标准物共扩增 ,计算出待测样本的个体的mRNA量。研究发现载脂蛋白E基因能在健康儿童周围血单核细胞表达 ,健康儿童载脂蛋白E基因表达量为 0 .37± 0 .15mol/molmRNA。表明采用竞争性逆转录—聚合酶链反应方法来检测人周围血单核细胞载脂蛋白E基因表达的分析方法快速、简便、灵敏、实用、可靠 ,且能准确定量 ,值得推广应用。 展开更多
关键词 分子生物学 载脂蛋白E基因表达的定量分析 逆转录—聚合酶链反应 竞争性 周围血单核细胞 儿童
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拟南芥CHS基因表达的实时荧光定量PCR检测 被引量:2
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作者 王艳 蒋磊 李韶山 《植物学通报》 CAS CSCD 北大核心 2005年第5期594-598,共5页
在简要介绍实时荧光定量PCR反应和定量原理的基础上,采用TaqMan荧光定量PCR技术,研究了UV-B辐射对拟南芥(Arabidopsisthaliana)CHS(查耳酮合成酶基因)表达的诱导,获得了与传统Northern杂交一致的结果。实时荧光定量PCR用于基因表达的定... 在简要介绍实时荧光定量PCR反应和定量原理的基础上,采用TaqMan荧光定量PCR技术,研究了UV-B辐射对拟南芥(Arabidopsisthaliana)CHS(查耳酮合成酶基因)表达的诱导,获得了与传统Northern杂交一致的结果。实时荧光定量PCR用于基因表达的定量检测,具有特异性强、自动化程度高、高效快捷,避免使用放射性同位素,能同时对多个样品中的起始模板进行准确定量等特点,因此该方法已逐渐被广泛用于基因表达的定量分析。 展开更多
关键词 拟南芥 查耳酮合成酶基因 基因表达定量分析 实时荧光定量PCR 荧光定量PCR检测 基因表达 NORTHERN杂交 CHS 放射性同位素 PCR反应
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实时荧光定量RT-PCR检测ICP4、IFN-γ和IL-18基因在ILTV感染鸡三叉神经节中的表达 被引量:2
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作者 木古丽 王云峰 +3 位作者 石星明 童光志 何来 王玫 《中国兽医学报》 CAS CSCD 北大核心 2008年第10期1150-1153,1166,共5页
将20只4月龄SPF鸡人工感染传染性喉气管炎病毒(ILTV)WG株,于感染后39d翅静脉和肌肉注射地塞米松(DEX),感染后5、9、10、11、12、39d及DEX处理后1、2、3、5、6、7、8d采喉拭子,检测病毒DNA,并于感染前和感染后6、10、20、31d及DEX处理后2... 将20只4月龄SPF鸡人工感染传染性喉气管炎病毒(ILTV)WG株,于感染后39d翅静脉和肌肉注射地塞米松(DEX),感染后5、9、10、11、12、39d及DEX处理后1、2、3、5、6、7、8d采喉拭子,检测病毒DNA,并于感染前和感染后6、10、20、31d及DEX处理后2、4、7、10、30d每组各取1只鸡颈静脉放血致死后无菌采集三叉神经节,提总RNA后利用real-timeRT-PCR的方法定量检测ICP4、IFN-γ和IL-18特异性mRNA。结果表明,在人工感染后处于潜伏状态的病毒能再被激活,并且ICP4基因在病毒还未被激活时就能低水平表达,这种低水平的立即早期(IE)基因的表达能提供持续的抗原刺激来维持神经节中的炎性细胞分泌IFN-γ和IL-18,以至于维持病毒处于潜伏状态而不致激活后感染其他的健康宿主;DEX处理后2d,IFN-γ的表达达到峰值,对病毒的再激活起抑制作用,尽管DEX处理前后IL-18的表达量没有明显变化,推测当潜伏的病毒一旦被激活时,IFN-γ可协同IL-18起到抑制IL-TV的作用。 展开更多
关键词 传染性喉气管炎病毒 地塞米松(DEX) 基因表达定量分析 实时荧光定量RT—PCR
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HIV及HSV感染妇女子宫阴道分泌物HIV-1及HSV-1/HSV-2的实时定量PCR分析
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《传染病网络动态》 2006年第3期13-13,共1页
据医学空间网2月8日报道(原载J Clin Microbiol.2006 Feb:44(2):423—32),有学者近期做了一项研究,对同时HIV及HSV感染的妇女的子宫阴道分泌物做了HIV-1 RNA和DNA及HSV—1/HSV-2 DNA的实时定量PCR检查。结果显示C(T)值的批... 据医学空间网2月8日报道(原载J Clin Microbiol.2006 Feb:44(2):423—32),有学者近期做了一项研究,对同时HIV及HSV感染的妇女的子宫阴道分泌物做了HIV-1 RNA和DNA及HSV—1/HSV-2 DNA的实时定量PCR检查。结果显示C(T)值的批内和批间变异系数为1.30%和0.69%(HIV-IRNA)及1.84%和0.67%(HIV-1 DNA)。HSV DNA的批内及批间变异系数为0.96%和1.49%。说明人工及自动方法从女性生殖器分泌物中联合提取HSV及HIV基因是可行的。总之,对于女性子宫阴道分泌物中HIV和HSV基因定量分析,实时定量PCR技术是独一无二的。 展开更多
关键词 实时定量PCR技术 HSV-2 HIV-1 阴道分泌物 感染妇女 PCR分析 HSV-1 子宫 女性生殖器 基因定量分析
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实时荧光定量PCR检测球孢白僵菌热休克蛋白基因hsp70在几种胁迫条件下的表达 被引量:9
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作者 谢翎 陈红梅 +3 位作者 汤强 蒲顺昌 李增智 黄勃 《菌物学报》 CAS CSCD 北大核心 2009年第6期806-812,共7页
本研究运用Realtime-PCR技术,对球孢白僵菌Beauveria bassiana hsp70基因在不同胁迫条件下的表达情况进行了检测。从mRNA转录水平探讨了不同胁迫条件对球孢白僵菌hsp70基因表达的影响。结果表明:38℃高温胁迫下,30min时表达量达到最高峰... 本研究运用Realtime-PCR技术,对球孢白僵菌Beauveria bassiana hsp70基因在不同胁迫条件下的表达情况进行了检测。从mRNA转录水平探讨了不同胁迫条件对球孢白僵菌hsp70基因表达的影响。结果表明:38℃高温胁迫下,30min时表达量达到最高峰,为对照样品的10.18倍。随后表达量开始下降,至180min时,其表达量降为最低,为对照样品的2.85倍;4℃低温胁迫下,2h检测到hsp70的表达量下降至最低点,为对照样品的0.25倍。随后表达量开始回升,至10h表达量始终维持在对照样品的1.4-1.5倍左右;紫外胁迫下(波长253.7nm),3min后hsp70的表达量快速上升至最高峰,为对照样品的2.33倍。随后表达量迅速下降,至60min表达量始终维持在对照样品的0.2倍左右。因此推测,hsp70基因在球孢白僵菌抵抗高温、低温和紫外三种胁迫方面都可能具有重要作用。同时研究结果也表明,球孢白僵菌hsp70基因启动子在逆境下可引导基因高效表达,因而在抗逆工程菌株构建方面可能具有重要的应用价值。 展开更多
关键词 昆虫病原真菌 基因表达定量分析 TAQMAN探针
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The quantified analysis of China's GM cotton yield capacity by C-D function and stochastic frontier model
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作者 张涛 薛宝娣 《Hunan Agricultural Science & Technology Newsletter》 2004年第1期11-13,共3页
Using a modified C D function and stochastic frontier model, the paper analyzed China's cotton yield capacity and found that the yield and technical efficiency of China's cotton planting system can be increas... Using a modified C D function and stochastic frontier model, the paper analyzed China's cotton yield capacity and found that the yield and technical efficiency of China's cotton planting system can be increased by the use of genetically modified (GM) varieties. 展开更多
关键词 GM cotton yield capacity C D function stochastic frontier model
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Quantitative analysis of a panel of gene expression in prostate cancer——with emphasis on NPY expression analysis 被引量:1
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作者 LIU Ai-jun FURUSATO Bungo +5 位作者 RAVINDRANATH Lakshmi CHEN Yong-mei SRIKANTAN Vasanta MCLEOD David G. PETROVICS Gyorgy SRIVASTAVA Shiv 《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》 SCIE CAS CSCD 2007年第12期853-859,共7页
Objective: To investigate molecular alterations associating with prostate carcinoma progression and potentially provide information toward more accurate prognosis/diagnosis. Methods: A set of laser captured microdis... Objective: To investigate molecular alterations associating with prostate carcinoma progression and potentially provide information toward more accurate prognosis/diagnosis. Methods: A set of laser captured microdissected (LCM) specimens from 300 prostate cancer (PCa) patients undergoing radical prostatectomy (RP) were defined. Ten patients representing "aggressive" PCa, and 10 representing "non-aggressive" PCa were selected based on prostate-specific antigen (PSA) recurrence, Gleason score, pathological stage and tumor cell differentiation, with matched patient age and race between the two groups. Normal and neoplastic prostate epithelial cells were collected with LCM from frozen tissue slides obtained from the RP specimens. The expressions of a panel of genes, including NPY, PTEN, AR, AMACR, DD3, and GSTP1, were measured by quantitative real-time RT-PCR (TaqMan), and correlation was analyzed with clinicopathological features. Results: The expressions of AMACR and DD3 were consistently up-regulated in cancer cells compared to benign prostate epithelial cells in all PCa patients, whereas GSTP1 expression was down regulated in each patient. NPY, PTEN and AR exhibited a striking difference in their expression patterns between aggressive and non-aggressive PCas (P=0.0203, 0.0284, and 0.0378, respectively, Wilcoxon rank sum test). The lower expression of NPY showed association with "aggressive" PCas based on a larger PCa patient cohort analysis (P=0.0037, univariate generalized linear model (GLM) analysis). Conclusion: Despite widely noted heterogeneous nature of PCa, gene expression alterations ofAM,4CR, DD3, and GSTP1 in LCM-derived PCa epithelial cells suggest for common underlying mechanisms in the initiation of PCa. Lower NPY expression level is significantly associated with more aggressive clinical behavior of PCa; PTEN and AR may have potential in defining PCa with aggressive clinical behavior. Studies along these lines have potential to define PCa-associated gene expression alterations and likely co-regulation of genes/pathways critical in the biology of PCa onset/progression. 展开更多
关键词 Prostate cancer NPY expression Quantitative real-time reverse-transcript polymerase chain reaction (RT-PCR)
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Overexpression of polo-like kinase1 predicts a poor prognosis in hepatocellular carcinoma patients 被引量:11
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作者 Zi-Li He He Zheng Hui Lin Xiong-Ying Miao De-Wu Zhong 《World Journal of Gastroenterology》 SCIE CAS CSCD 2009年第33期4177-4182,共6页
AIM: To elucidate the role of overexpressed polo-like kinasel (PLK1)in hepatocellular carcinoma (HCC). METHODS: We prospectively collected clinicopathological, immunohistochemical and semi-quantitative reverse t... AIM: To elucidate the role of overexpressed polo-like kinasel (PLK1)in hepatocellular carcinoma (HCC). METHODS: We prospectively collected clinicopathological, immunohistochemical and semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) data from 135 HCC patients undergoing successful hepatectomy. The correlations between PLK1 mRNA expression and clinicopathologic variables were analyzed by Mann-Whitney U test. Prognostic factors were identified by univariate and multivariate analyses. RESULTS: Immunohistochemical results showed overexpression of PLK1 was mainly found in tumor tissues compared with tumor-free tissue. A similar mRNA result was obtained by semi-quantitative RT-PCR. A total of 111 samples were positive for PLK1 mRNA expression. The positive expression was correlated with venous invasion, tumor nodules and Edmondson grade. Furthermore, 1, 3, 5-year survival rates in the positive expression group were significantly lower than the negative control group. Multivariate analysis showed that positive PLK1 expression was an independent risk factor for HCC. CONCLUSION: PLK1 could be a potential biomarker for diagnosis and therapy for HCC. 展开更多
关键词 Hepatocellular carcinoma IMMUNOHISTOCHEMISTRY Reverse transcription-polymerase chain reaction Survival analysis Polo-like kinase 1
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Identification and expression analysis of bovine ANGPTL gene family 被引量:2
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作者 Zhengbing GUAN Guolin CAI +1 位作者 Junyong SUN Jian LU 《Current Zoology》 SCIE CAS CSCD 北大核心 2010年第4期445-453,共9页
Encoded by seven genes, angiopoietin-like (ANGPTL) family members structurally similar to the angiogenic regulating factor angiopoietin are known to possess biological activities in angiogenesis and metabolism. Here... Encoded by seven genes, angiopoietin-like (ANGPTL) family members structurally similar to the angiogenic regulating factor angiopoietin are known to possess biological activities in angiogenesis and metabolism. Here we reports for the first time the identification and expression analysis of all the seven members of bovine ANGPTL gene family, which were designated bANGPTL1 to bANGPTL7 in order. The seven bANGPTL genes consist of 4-9 exons, span 3800M-3000 bp and are located on different chromosomes. The deduced amino acid sequences of the members all possess an N-terminal coiled-coil domain and a C-terminal fibrinogen-like domain, both characteristics of angiopoietins. Phylogenetic analysis showed that the 32 identified ANGPTL homologs from 9 species could be classified into two major groups. Real-time quantitative PCR (Q-PCR) analysis revealed that the bANGPTL family members have different expression patterns. This study will be helpful for investigation on the biological role of the bANGPTL family in this economically important species. Furthermore, it provides an insight into the molecular evolution of the emerging ANGPTL family 展开更多
关键词 CATTLE Angiopoietin-like Phylogenetic analysis Q-PCR
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Expression and semi-quantification of hepatitis B virus reverse transcriptase protein in a prokaryotic system
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作者 CHAO ZHAO YONG XIANG WANG ZHENG HONG YUAN YU MEI WEN 《Journal of Microbiology and Immunology》 2006年第3期189-193,共5页
The reverse transcriptase (RT) protein of hepatitis B virus (HBV) has been successfully expressed by recombinant technology in Eschericahia coli ( E. coli ). In this study we aimed to develop a semi-quantitative... The reverse transcriptase (RT) protein of hepatitis B virus (HBV) has been successfully expressed by recombinant technology in Eschericahia coli ( E. coli ). In this study we aimed to develop a semi-quantitative assay for the study of HBV RT protein using this system. Complete HBV polymerase gene from a wild type virus (rt306P) and the polymerase gene from a mutant, with rt306P substituted by serine (rtP306S) were separately fused to the maltose binding protein (MBP) gene and expressed in E. coli respectively. The expression levels of HBV polymerase genes from the wild type virus and its counterpart mutant at rt306 were compared. When these proteins were semi-quantified by Westem blotting using rabbit anti-TP serum, the rtP306S mutant showed decreased expression of MBP-HBV polymerase. By this method, we have shown that the expression level of HBV RT could be affected by substitutions in its amino acid sequences, and this method could be used to study the characteristics of HBV RT protein. 展开更多
关键词 Hepatitis B virus DNA polymerase expression Mutants
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Expression Analysis of the ANS Gene in Fragaria x ananassa cv, Toyonaka
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作者 Xiaonan Zhang Qing Chen Haowei Yu Shuli Zhou Haoru Tang 《Journal of Life Sciences》 2012年第11期1185-1189,共5页
Strawberry is an economically valuable crop all over the word. Its fruits contain large amounts of polyphenol compounds, anthocyanins for instance. Anthocyanins play important roles in attracting pollinators and prote... Strawberry is an economically valuable crop all over the word. Its fruits contain large amounts of polyphenol compounds, anthocyanins for instance. Anthocyanins play important roles in attracting pollinators and protecting plants from various stresses. ANS (Anthocyanidin synthase) catalyzes the conversion of leucoanthocyanidins to colored anthocyanidins. In this study, the total anthocyanins concentration at seven developmental stages of the strawberry fruit were investigated by UV spectrophotometry at 496 nm and 700 rim; and the expression levels of the Fa-ANS gene were studied by qRT-PCR. The results showed that the anthocyanins contents gradually increased along with the fruit maturation; while the expression patterns of Fa-ANS were consistent with the anthocyanins. 展开更多
关键词 Anthocyanins ANS strawberry fruit developmental stages.
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番茄早疫病菌Hog1 MAPK同源基因AsHog1的表达特性及其与抗药性的关联性 被引量:7
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作者 任璐 刘慧平 韩巨才 《菌物学报》 CAS CSCD 北大核心 2010年第4期542-547,共6页
根据几种丝状真菌Hog1 MAPK(丝裂原活化蛋白激酶)的保守氨基酸序列设计简并引物,从番茄早疫病菌Alternaria solani中扩增出MAPK同源基因的部分片段,命名为AsHog1。用Real-timeRT-PCR技术比较了AsHog1在异菌脲敏感菌株和抗性菌株中的表... 根据几种丝状真菌Hog1 MAPK(丝裂原活化蛋白激酶)的保守氨基酸序列设计简并引物,从番茄早疫病菌Alternaria solani中扩增出MAPK同源基因的部分片段,命名为AsHog1。用Real-timeRT-PCR技术比较了AsHog1在异菌脲敏感菌株和抗性菌株中的表达特性。结果表明,敏感菌株QX21经0.8mol/L NaCl和2mg/L异菌脲处理后,AsHog1相对表达量持续升高,在12h达到最大值,分别为对照的4.12倍和25.23倍,16h略有降低;而室内诱导抗性突变体和田间抗性菌株经相同处理后的AsHog1表达量变化不大,且明显低于敏感菌株。由此推测,番茄早疫病菌AsHog1基因表达特征与其对异菌脲抗药性相关。研究结果为深入研究病原菌对异菌脲抗药性的分子机理、建立抗药性分子监测技术、延缓和防止抗药性的产生及发展奠定了一定的理论基础。 展开更多
关键词 番茄早疫病菌 抗药性机理 基因表达定量分析 丝裂原活化蛋白激酶(MAPK) 异菌脲
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