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大肠杆菌trpBA和serA基因的串联表达 被引量:4
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作者 张绪梅 郭长江 +1 位作者 刘云 徐琪寿 《微生物学通报》 CAS CSCD 北大核心 2009年第1期2-8,共7页
大肠杆菌trpBA基因编码的色氨酸合成酶(tryptophan synthetase,TSase)是色氨酸合成的关键酶;serA基因编码的磷酸甘油酸脱氢酶(D-3-phosphoglycerate-dehydrogenase,PGDH)为L-丝氨酸合成(色氨酸合成的底物)的关键酶。为了通过... 大肠杆菌trpBA基因编码的色氨酸合成酶(tryptophan synthetase,TSase)是色氨酸合成的关键酶;serA基因编码的磷酸甘油酸脱氢酶(D-3-phosphoglycerate-dehydrogenase,PGDH)为L-丝氨酸合成(色氨酸合成的底物)的关键酶。为了通过基因工程手段来增加色氨酸的产量,在利用高效的原核表达载体pET22b(+)分别对trpBA和serA基因克隆表达的基础上,采用PCR方法扩增了抗反馈抑制的serA和trpBA基因,将两基因串联于pET22b(+)载体上,共构建了4种方式的串联质粒,实现了2种蛋白酶在大肠杆菌中的共表达。聚丙烯酰胺电泳分析显示,ABA—I重组菌株在37kD(PGDH)、29kD(色氨酸合成酶的α,亚基)、44kD(β亚基)处均有明显的蛋白表达带。4种串联表达质粒重组菌的TSase酶活性,分别比含空载体菌相应酶的活性提高2~4倍,PGDH酶活性分别提高约2.1~3.6倍。经摇瓶发酵实验表明酶活性较高的ABA—I菌株色氨酸合成量亦最高,约为对照菌株的20.2倍。 展开更多
关键词 色氨酸 大肠杆菌 sera基因 trpBA基因 串联表达
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L-丝氨酸产生菌c-11中serA基因的克隆与表达 被引量:1
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作者 谢玉清 罗淑萍 茆军 《新疆农业科学》 CAS CSCD 2007年第5期652-653,共2页
利用PCR技术以L-丝氨酸产生菌株黄色短杆菌c-11的染色体DNA为摸板,扩增出serA基因,连接到表达载体pEC7上,转化L-丝氨酸产生菌c-11,使其氨基酸产量由12 g/L增加到18 g/L,产酸率增加了50%。
关键词 黄色短杆菌C-11 sera基因 克隆 表达
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抗恶性疟SERA噬菌体抗体库的构建及单链抗体的筛选与鉴定(英文)
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作者 徐伟文 李文全 +2 位作者 李明 毕惠祥 Toshihiro Horii 《细胞与分子免疫学杂志》 CAS CSCD 北大核心 2002年第1期58-62,共5页
目的 研制用于疟疾简便、快速诊断试剂盒的试剂。方法 利用噬菌体抗体库技术,构建抗恶性疟原虫丝氨酸重复抗原(SERA)的噬菌体抗体库。经3轮“吸附-洗脱-扩增”的富集反应后,从中筛选抗SERA的阳性克隆株,并进行可溶性诱... 目的 研制用于疟疾简便、快速诊断试剂盒的试剂。方法 利用噬菌体抗体库技术,构建抗恶性疟原虫丝氨酸重复抗原(SERA)的噬菌体抗体库。经3轮“吸附-洗脱-扩增”的富集反应后,从中筛选抗SERA的阳性克隆株,并进行可溶性诱导表达,最后用ELISA和Western blot等进行鉴定。结果 成功地构建了中等库容的噬菌体抗体库,并从中筛选到9株抗SERA的阳性克隆株,表达的单链抗体的相对分子质量大小约为31kDa,能与 SE47'抗原起特异性结合反应。结论 成功地构建了抗SE47'的噬菌体抗体库,从中筛选制备的单链抗体为研制疟疾快速诊断试剂盒奠定了基础。 展开更多
关键词 噬菌体抗体库 单链抗体 丝氨酸重复抗原 sera 恶性疟
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Effects of sera from burn patients on human hepatocytic viscoelasticity 被引量:2
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作者 WANG Xiao Jun, LUO Xiang Dong, LUO Qin and YANG Zong Cheng Burn Research Institute, Southwest Hospital, Third Military Medical University, Chongqing 400038, China 《World Journal of Gastroenterology》 SCIE CAS CSCD 1998年第1期65-65,共1页
EfectsofserafromburnpatientsonhumanhepatocyticviscoelasticityWANGXiaoJun,LUOXiangDong,LUOQinandYANGZongCh... EfectsofserafromburnpatientsonhumanhepatocyticviscoelasticityWANGXiaoJun,LUOXiangDong,LUOQinandYANGZongChengBurnResearchIn... 展开更多
关键词 burns/pathology IMMUNE sera liver/pathology
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Expressions of ICAM-1 and its mRNA in sera and tissues of patients with hepatocellular carcinoma 被引量:14
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作者 Jing Xu1 Ming Hui Mei1 +3 位作者 Si En Zeng2 Qing Fen Shi3 Yong Ming Liu4 Li Ling Qin3 1Department of Hepatobiliary Surgery2Department of Pathology3Institute of Hepatobiliary Surgery4Department of Biochemistry, Guilin 541001, Guangxi Province, China 《World Journal of Gastroenterology》 SCIE CAS CSCD 2001年第1期120-125,共6页
INTRODUCTIONThe increased expression of ICAM-1 on a widerange of cells and in the sera of patients withmalignancies, chronic liver diseases andinflammation diseases has been described since thelate 1980s[1-22]. Recent... INTRODUCTIONThe increased expression of ICAM-1 on a widerange of cells and in the sera of patients withmalignancies, chronic liver diseases andinflammation diseases has been described since thelate 1980s[1-22]. Recently rapid progress in studieson expression of ICAM-1 in patients withhepatocellular carcinoma ( HCC ) have beenachieved, including clinical and experimentalresearches[23-31]. 展开更多
关键词 LIVER neoplasms/diagnosis intercellular adhesion molecule-1 RNA messenger alpha-fetoprotein immunohistochemistry polymerase chain reaction radioimmunoassay CLC Number:R57 Document ID:A INTRODUCTIONThe increased EXPRESSION of ICAM-1 on a widerange of cells and in the sera of PATIENTS withmalignancies chronic LIVER DISEASES andinflammation DISEASES has been described since thelate 1980s[1-22]. Recently rapid progress in studieson EXPRESSION of ICAM-1 in PATIENTS withhepatocellular carcinoma ( HCC ) have beenachieved including clinical and experimentalresearches[23-31].……
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Get effective polyclonal antisera in one month 被引量:45
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作者 YUAN XIN HU, Ju YUAN QUO, Lu SHEN, YAN CHEN, Zu CHUAN ZHANG, YONG LIAN ZHANGState Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 320 Yue Yang Road, 《Cell Research》 SCIE CAS CSCD 2002年第2期157-160,共4页
According to the traditional immunization procedure, after the first injection of the sample A (emulsion of aimed antigen and Freund's complete adjuvant) to immunize rabbit, successive injections of the sample B (... According to the traditional immunization procedure, after the first injection of the sample A (emulsion of aimed antigen and Freund's complete adjuvant) to immunize rabbit, successive injections of the sample B (emulsion of aimed antigen and Freund's incomplete adjuvant) were followed every 2-4 weeks. In general,high titer of the corresponding polyclonal antisera will be observed after 4-5 injections of sample B in 3-4months. This report presents a simply modified procedure that was able to stimulate the antisera formation in one month and achieve enough avidity to satisfy either Western blot or immunohistochemistry analysis.It just applied an additional injection of the sample A to the rabbit at the 3rd day after the primary immunization injection. You could gain the high titer of the antisera right after the first sample B injection in one month. This method has produced the desired results in three different recombinant antigens with different molecular weight (5.9 KD-55 KD) expressed from prokaryotic or eukaryotic cells. 展开更多
关键词 POLYCLONAL ANTIsera preparation Modified IMMUNIZATION method.
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Comparative Study on 4 EIA Kits for Screening Antibody to Hepatitis C Virus in Pooled Sera
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作者 LIU PEI SHI ZHI-XU +3 位作者 ZHANG YAN-CHAO AND ZHANG XIN-YING (Deportment of Public Health, twrtment of Infectious Disease,Xuzhou Medical College Xuzhou Red Cross BloodService, Xuzhou, Jiangsu, China) 《Biomedical and Environmental Sciences》 SCIE CAS CSCD 1998年第1期75-80,共6页
Four enzyme immunoassay (EIA) test kits, 1 Canadian product and 3 Chinese products,were used in the comparative study. Each pool consisted of 5 sera, and the 5 single sera were tested as controls. The tests were carri... Four enzyme immunoassay (EIA) test kits, 1 Canadian product and 3 Chinese products,were used in the comparative study. Each pool consisted of 5 sera, and the 5 single sera were tested as controls. The tests were carried out according to the instructions, keeping the same dilution of each serum in single and pool samples. It was found that with the Canadian kit,the positive and negative results of opled sera had no difference from that of the controls (P>0. 10). In the case of Chinese Yali and Kehua kits, the positive results of pooled sera showed no difference from the controls (P >0. 10), but the optical density (OD) of negative opls were increased (P < 0. 01 ), though quite distant from the cut-off values. In the case of Changzheng kit, the OD of opitive opls were significantly lower than those of the controls (P < 0. 05 ), and weak positive samples missed the detection. However this problem could be overcome by blocking the microwells beforehand. Our experiment demonstrate that not all EIA test kits are suitable for screening opls for antithey to hepatitis C virus, and that it is important to assess the sensitivity of the EIA kit to be used for this purpose. 展开更多
关键词 EIA Comparative Study on 4 EIA Kits for Screening Antibody to Hepatitis C Virus in Pooled sera
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Role of Soluble Factors in Immune and Normal Sera in the Control of Giardiasis Due to Giardia Iamblia
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作者 R.K.SRIVASTAVA N.SETHI +1 位作者 N.GUPTA A.K.AGARWAL 《Biomedical and Environmental Sciences》 SCIE CAS CSCD 1992年第2期87-91,共5页
Hyperimmune sera (HIS), raised against crude giardia antigen, on in vitro interaction, caused more agglutination of Giardia lamblia trophozoites. Heat inactivated HIS possessed a comparable agglutinating activity as t... Hyperimmune sera (HIS), raised against crude giardia antigen, on in vitro interaction, caused more agglutination of Giardia lamblia trophozoites. Heat inactivated HIS possessed a comparable agglutinating activity as the non-inactivated controls. Non-inactivated normal (unimmunized) serum caused immobilization of Giardia trophozoite, which was checked on heat inactivation. Antibodies in immune sera are mainly responsible for agglutination, whereas the heat labile non-immune components control the mobility of the intestinal parasite. 展开更多
关键词 Role of Soluble Factors in Immune and Normal sera in the Control of Giardiasis Due to Giardia Iamblia
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用SERA技术检测线路板锡涂层的性能
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《印制电路与贴装》 2001年第6期31-33,27,共4页
关键词 sera技术 线路板 锡涂层 检测 性能 印制电路
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SERA肽类似物具抗疟疾的保护作用
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作者 陈锦荣 江丽君 《生物制品快讯》 2004年第8期6-7,共2页
关键词 sera肽类似物 抗疟疾 保护作用 疫苗
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MIL—P—55110E附录连续电化学还原分析(SERA)可焊性试验
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《印制电路信息》 1996年第9期38-43,共6页
1.范围1.1 范围 本试验方法介绍了可焊接表面的氧化等级评定方法和程序。铜、锡和铅表面的氧化物种类和数量,对可焊性有显著的影响。本方法需要用电化学还原技术,来确定电镀通孔、焊接盘及印制线路板表面导线上的氧化物类型和数量。SER... 1.范围1.1 范围 本试验方法介绍了可焊接表面的氧化等级评定方法和程序。铜、锡和铅表面的氧化物种类和数量,对可焊性有显著的影响。本方法需要用电化学还原技术,来确定电镀通孔、焊接盘及印制线路板表面导线上的氧化物类型和数量。SERA可焊性试验方法,可取代本规范要求的其他可焊性试验方法。 展开更多
关键词 电化学还原 印制线路板 可焊性试验 测试头 镀通孔 sera 参考电极 试验方法 储液器 可编程电源
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Sera Monastery's Home for the Elderly
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作者 SUO QIONG 《The Journal of Human Rights》 2007年第1期32-32,共1页
Xirao Jiangcan, a 78-year- old lama, is sunbathing in front of a scripture-chanting hall in Sera Monastery in Lhasa, Tibet. He feels comfortable in the native of Deqin warmth of the sun. A County in Diqing Tibetan Aut... Xirao Jiangcan, a 78-year- old lama, is sunbathing in front of a scripture-chanting hall in Sera Monastery in Lhasa, Tibet. He feels comfortable in the native of Deqin warmth of the sun. A County in Diqing Tibetan Autonomous Prefecture, Yunnan, Xirao came to Lhasa at the age of 26. He has lived in Sera Monastery for 52 years. Currently he resides in the monastery's Old People's Home. Many other old lamas also live in the old people's home. 展开更多
关键词 HOME sera Monastery’s Home for the Elderly
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The Vicissitudes of the Sera Monastery
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《China's Tibet》 1995年第2期6-9,共4页
TheVicissitudesoftheSeraMonastery¥GARDAWACERINGApanoramicviewoftheSeraMonastery.TheSeraMonasteryWhichliessom... TheVicissitudesoftheSeraMonastery¥GARDAWACERINGApanoramicviewoftheSeraMonastery.TheSeraMonasteryWhichliessomefivekmnorthofLha... 展开更多
关键词 The Vicissitudes of the sera Monastery
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From Geshi of Sera Monastery to KMT Attache
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作者 Xerab Nyima 《China's Tibet》 1995年第1期20-22,共3页
FromGeshiofSeraMonasterytoKMTAttachebyXerabNyimaThedeafeningsoundofdrumsandmusicalinstrumentsresoundedinGyai... FromGeshiofSeraMonasterytoKMTAttachebyXerabNyimaThedeafeningsoundofdrumsandmusicalinstrumentsresoundedinGyaincain—OfficialGre... 展开更多
关键词 From Geshi of sera Monastery to KMT Attache
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Distinctive serotypes of SARS-related coronaviruses defined by convalescent sera from unvaccinated individuals 被引量:3
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作者 Chee Wah Tan Feng Zhu +17 位作者 Wan Ni Chia #Barnaby EYoung Aileen Ying Yan Yeoh Thomas Althaus Chee Fu Yung Wee Chee Yap Beng Lee Lim Mark I-C Chen Jinyan Zhang Yun Yan Mah Eric Voiglio Alex Sigal Jianxin Huo Shengli Xu Yee Joo Tan Kong-Peng Lam David Lye Lin-Fa Wang 《hLife》 2023年第1期26-34,共9页
Multiple Omicron sub-lineages have emerged,with Omicron XBB and XBB.1.5 subvariants becoming the dominant variants globally at the time of this study.The key feature of new variants is their ability to escape humoral ... Multiple Omicron sub-lineages have emerged,with Omicron XBB and XBB.1.5 subvariants becoming the dominant variants globally at the time of this study.The key feature of new variants is their ability to escape humoral immunity despite the fact that there are limited genetic changes from their preceding variants.This raises the question of whether Omicron should be regarded as a separate serotype from viruses serologically clustered with the ancestral severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)virus.Here,we present cross-neutralization data based on a pseudovirus neutralization test using convalescent sera from naïve individuals who had recovered from primary infection by SARS-CoV-1 and SARS-CoV-2 strains/variants including the ancestral virus and variants Beta,Delta,Omicron BA.1,Omicron BA.2 and Omicron BA.5.The results revealed no significant cross-neutralization in any of the three-way testing for SARS-CoV-1,ancestral SARS-CoV-2 and SARS-CoV-2 Omicron subvariants.The data argue for the assignment of three distinct serotypes for the currently known human-infecting SARS-related coronaviruses. 展开更多
关键词 SARS-CoV-2 SARS-CoV-1 SEROTYPE Omicron primary infection convalescent sera
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甲基营养菌M.sp.MB200中serA基因功能初探 被引量:1
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作者 蒋琼 马晟 +5 位作者 周侃 李献 唐咸来 李俊芳 张敏 申佩弘 《基因组学与应用生物学》 CAS CSCD 北大核心 2014年第3期551-555,共5页
serA基因编码3-磷酸甘油酸脱氢酶(PGDH),此酶在合成L-丝氨酸的第一步起催化作用。利用质粒pCM80分别与野生型基因serA、缺失了C末端ACT功能域的突变基因serA△77构建了重组体表达菌MB200/pCM80serA和MB200/pCM80serA△77。通过分析发现... serA基因编码3-磷酸甘油酸脱氢酶(PGDH),此酶在合成L-丝氨酸的第一步起催化作用。利用质粒pCM80分别与野生型基因serA、缺失了C末端ACT功能域的突变基因serA△77构建了重组体表达菌MB200/pCM80serA和MB200/pCM80serA△77。通过分析发现野生型基因重组表达菌的PGDH酶活受L-丝氨酸浓度影响较大,当反应体系中的L-丝氨酸浓度为40μmol/L时,酶活减少了约1/3;缺失了ACT功能域的基因表达菌的PGDH酶活基本不变,不受L-丝氨酸浓度的影响。进一步在静息细胞反应条件下,对重组菌和野生菌株MB200进行发酵产L-丝氨酸的研究,实验发现MB200/pCM80serA产L-丝氨酸的量为13.6mg/mL,MB200/pCM80serA△77产L-丝氨酸的量为16.8 mg/mL,而野生型菌株M.sp MB200的产量为6.4 mg/mL,重组菌的L-丝氨酸产量都有所提高,且MB200/pCM80serA△77的产量提高更多。结果证实了甲基营养菌MB200中serA基因与产L-丝氨酸具有反馈抑制关系,为进一步解除反馈抑制并发酵产L-丝氨酸奠定了基础。 展开更多
关键词 甲基营养菌MB200 sera L-丝氨酸 反馈抑制
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Safety of convalescent sera for the treatment of viral severe acute respiratory syndrome: an experimental model in rhesus macaque 被引量:1
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作者 HE Wei-ping LI Bo-an +1 位作者 ZHAO Jun CHENG Yun 《Chinese Medical Journal》 SCIE CAS CSCD 2013年第19期3790-3792,共3页
Serotherapy has assumed different clinical roles over the years.With the development of antibiotics,the use of serotherapy to treat bacterial diseases declined.However,interest in viral serotherapy increased due to th... Serotherapy has assumed different clinical roles over the years.With the development of antibiotics,the use of serotherapy to treat bacterial diseases declined.However,interest in viral serotherapy increased due to the lack of a specific therapy for viral infections.Human convalescent sera have been used to treat and prevent measles,mumps,polio,Spanish flu,vaccinia and varicella viral infection,among others. 展开更多
关键词 severe acute respiratory syndrome convalescent sera SAFETY
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Identification of new centrosome proteins by autoimmune patient sera
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作者 XIA Liang1, LI Yan1, YANG Dong1, WANG LiMin1, HE Fang1, ZHOU ChunYuan1, LI YongZhe2, ZENG ChangQing1 & He DaCheng1 1 Key Laboratory for Cell Proliferation and Regulation Biology of Ministry of Education, Beijing Normal University, Beijing 100875, China 2 Peking Union Medical Hospital, Peking Union Medical College, Beijing 100730, China 3 Beijing Genomic Institute, Beijing 101300, China 《Science China(Life Sciences)》 SCIE CAS 2007年第2期194-202,共9页
Compared to other subcellular organelles, centrosome proteome can hardly be studied, due to the dif- ficulties in separation and purification of centrosome. Auto-antisera from 6 autoimmune patients, which recognized c... Compared to other subcellular organelles, centrosome proteome can hardly be studied, due to the dif- ficulties in separation and purification of centrosome. Auto-antisera from 6 autoimmune patients, which recognized centrosome specifically in immunofluorescence, were used to identify the corresponding centrosomal proteins. The sera were first tested by Western blot on whole cell lysate, and all bound antibodies were then eluted from each single band in Western blot membrane to assure which antibody was responsible for the centrosome specific immunofluorescence staining. The corresponding pro- teins were obtained by immunoprecipitation and identified by mass spectrometry. Six centrosomal proteins, including 2 known centrosomal proteins and 4 proteins with unknown localization or report- edly non-centrosomal localization, were identified. These proteins apparently involve in cell cycle regulation, signal transduction pathways, molecular chaperons, and metabolism enzymes, which may reflect the expected functional diversity of centrosome. 展开更多
关键词 CENTROSOME AUTOIMMUNE PATIENT sera ELUTION of antibody immunoflourescence IMMUNOPRECIPITATION mass spectrometry
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Application of Conglutinin-ELISA for the detection of Circulating Immune Complexes in Sera of Patients with Epidemic Hemorrhagic Fever.
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《Chinese Medical Journal》 SCIE CAS CSCD 1994年第7期57-57,共1页
Conglutinin was extracted and purified from bovine’s sera and was used in ELISA for the detection of circulating immune complexes in the sera of patients suffering from epidemic hemorrhagic fever (EHF). The detected ... Conglutinin was extracted and purified from bovine’s sera and was used in ELISA for the detection of circulating immune complexes in the sera of patients suffering from epidemic hemorrhagic fever (EHF). The detected rates of circu- 展开更多
关键词 ELISA EHF Application of Conglutinin-ELISA for the detection of Circulating Immune Complexes in sera of Patients with Epidemic Hemorrhagic Fever
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Protein array of Coxiella burnetii probed with Q fever sera 被引量:2
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作者 WANG XiLe XIONG XiaoLu +2 位作者 GRAVES Stephen STENOS John WEN BoHai 《Science China(Life Sciences)》 SCIE CAS 2013年第5期453-459,共7页
Coxiella burnetii is the etiological agent of Q fever.To identify its major seroreactive proteins,a subgenomic protein array was developed.A total of 101 assumed virulence-associated recombinant proteins of C.burnetii... Coxiella burnetii is the etiological agent of Q fever.To identify its major seroreactive proteins,a subgenomic protein array was developed.A total of 101 assumed virulence-associated recombinant proteins of C.burnetii were probed with sera from mice experimentally infected with C.burnetii and sera from Q fever patients.Sixteen proteins were recognized as major seroreactive antigens by the mouse sera.Seven of these 16 proteins reacted positively with at least 45% of Q fever patient sera.Notably,HspB had the highest fluorescence intensity value and positive frequency of all the proteins on the array when probed with both Q fever patient sera and mouse sera.These results suggest that these seven major seroreactive proteins,particularly HspB,are potential serodiagnostic and subunit vaccine antigens of Q fever. 展开更多
关键词 阵列探测 蛋白质 热毒 人血清 亚单位疫苗 血清学诊断 实验感染 重组蛋白
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