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苹果褪绿叶斑病毒外壳蛋白基因的原核表达及抗血清制备 被引量:3
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作者 耿超 牟安丽 +2 位作者 刘志强 李向东 周涛 《果树学报》 CAS CSCD 北大核心 2012年第5期755-758,I0002,共5页
【目的】为了制备苹果褪绿叶斑病毒(Apple chlorotic leaf spot virus,ACLSV)的抗血清,【方法】将ACLSV外壳蛋白(Coat protein,CP)基因克隆到原核表达载体pEHISTEV和pEHISEGFPTEV,转化大肠杆菌Rosetta,切胶回收诱导表达的CP融合蛋白免... 【目的】为了制备苹果褪绿叶斑病毒(Apple chlorotic leaf spot virus,ACLSV)的抗血清,【方法】将ACLSV外壳蛋白(Coat protein,CP)基因克隆到原核表达载体pEHISTEV和pEHISEGFPTEV,转化大肠杆菌Rosetta,切胶回收诱导表达的CP融合蛋白免疫新西兰长耳兔,制备ACLSV的抗血清。【结果】利用原核表达载体pEHISTEV和pE-HISEGFPTEV在Rosetta中分别表达出了分子质量为25 ku和55 ku的ACLSV CP融合蛋白。切胶回收25 ku的融合蛋白免疫新西兰长耳兔。经ELISA测定,所制备的抗血清效价为1∶1 024,Western blot及组织印迹分析表明抗血清能与ACLSV CP发生特异性的免疫反应。【结论】该血清特异性强,效价高,可用于苹果叶片、果实和枝条等样品中A-CLSV的检测。 展开更多
关键词 苹果褪绿叶斑病毒 外壳蛋白基因 原核表达 抗血清 组织印迹分析
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Decreased expression of Neurensin-2 correlates with poor prognosis in hepatocellular carcinoma 被引量:7
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作者 Hai-Qing Ma Xiao-Ting Liang Jing-Jing Zhao Hui Wang Jian-Cong Sun Yi-Bing Chen Ke Pan Jian-Chuan Xia 《World Journal of Gastroenterology》 SCIE CAS CSCD 2009年第38期4844-4848,共5页
AIM: To investigate the expression of Neurensin-2 (NRSN2) in hepatocellular carcinoma (HCC) and its prognostic values in predicting survival. METHODS: The expression and prognostic significance of NRSN2 in HCC w... AIM: To investigate the expression of Neurensin-2 (NRSN2) in hepatocellular carcinoma (HCC) and its prognostic values in predicting survival. METHODS: The expression and prognostic significance of NRSN2 in HCC was examined by performing immunohistochemical analysis using a total of 110 HCC clinical tissue samples, and Western blotting analysis to further confirm the result. RESULTS: Decreased NRSN2 expression was shown in 70.9% cases. Loss of NRSN2 expression in HCC was significantly related to tumor size (P = 0.006). Larger tumor size was related to negative expression of NRSN2. Patients showing negative NRSN2 expression had a significantly shorter overall survival than those with positive expression (P = 0.008). Multivariate Cox regression analysis indicated that NRSN2 expression level was an independent factor of survival (P = 0.013). Western blotting analysis further confirmed decreased expression of NRSN2 in tumor tissues compared with non-tumorous tissues. CONCLUSION: Our study indicated that NRSIV2 could be a tumor suppressor gene for HCC and a candidate biomarker for long-term survival in HCC. 展开更多
关键词 Neurensin-2 Hepatocellular carcinoma EXPRESSION Immunohistochemical analysis PROGNOSIS
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Expression of PADI4 in hepatocellular carcinoma 被引量:1
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作者 Yang Lv Yan Xia Yaohua Wang Chongyuan Cai 《The Chinese-German Journal of Clinical Oncology》 CAS 2009年第8期453-455,共3页
Objective: The aim of the research was to study peptidylarginine deiminase type 4 (PAD4/PADI4) expression and its tumodgenic mechanism in hepatocellular carcinomas. Methods: Expressions of PADI4 and p53 were inves... Objective: The aim of the research was to study peptidylarginine deiminase type 4 (PAD4/PADI4) expression and its tumodgenic mechanism in hepatocellular carcinomas. Methods: Expressions of PADI4 and p53 were investigated in tumors and non-tumor tissues by Western blot in patients with hepatocellular carcinomas. We constructed plasmid of PADI4-Flag and transfected it in Hela cells to investigate the mechanism. Results: Western blot analysis showed higher PADI4 expression in hepatocellular carcinomas than in the surrounding healthy tissues. Furthermore, by Western blot, we detected decreased p53 levels in the tumor tissues of patients with hepatocellular carcinomas compared to surrounding healthy tissues. In Hela cells transfected with PcDNA3.0-Flag-PADI4 plasmid, the expression of p53 decreased obviously. Conclusion: Our results suggest that PADI4 elevated in the tissues of hepatocellular carcinomas and induced tumorigenic by down-regulating p53 expression. 展开更多
关键词 hepatocellular carcinoma peptidylarginine deiminase type 4 (PAD4/PADI4) tumorigenic mechanism
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Murine brain tissues with human cytomegalovirus infection: a proteomic study
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作者 Xie Ni Yuan Jianhui +1 位作者 Cai Yinsha Liu Jianjun 《Journal of Medical Colleges of PLA(China)》 CAS 2010年第2期65-74,共10页
To establish two-dimensional electrophoresis profiles with high resolution and reproducibility from murine brain tissues by human cytomegalovirus(HCMV) infection and paired murine brain tissues and to identify the d... To establish two-dimensional electrophoresis profiles with high resolution and reproducibility from murine brain tissues by human cytomegalovirus(HCMV) infection and paired murine brain tissues and to identify the differential expression proteins. Methods: Forty Kunming mice were randomly divided into infection group (20) injected with HCMVAD169 and control group (20) injected with saline into their brain. After 30 days, the murine brain tissues by HCMV infection and paired murine brain tissues were separated by two-dimensional gel electrophoresis(2-DE), analyzed by Image Master 2D software, and identified by peptide mass fingerprint(PMF) and database searching, and make Western blotting analyses the differential expression of individual proteins. Results: Well resolved, reproducible 2-D maps of the above tissues were obtained. Some of the different proteins identified by mass spectrometry(MS) were matched in the SWISS-2D PAGE database, Western blotting analyses were further carried out to verify the differential expression of individual proteins. Conclusion: These data will be valuable for studying the diagnosis of disease at an early stage, mechanisms of pathogenic and the key to the development of anti-HCMV medicine. 展开更多
关键词 Human cytomegalovirus INFECTION PROTEOMICS Differential expression
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