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鸡IL-2基因的克隆及序列测定 被引量:15
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作者 陈奖励 王欢 +4 位作者 宋英晖 宋秀龙 陈红岩 姜永厚 卢景良 《中国兽医杂志》 CAS 北大核心 2000年第4期11-13,共3页
应用逆转录多聚酶链反应 (RT- PCR)技术 ,参照 Sundick发表的鸡白细胞介素 2 (IL- 2 ) c DNA基因序列 ,利用自行设计合成的 1对引物 ,从 Con A活化的 5周龄 HWL - SPF鸡脾细胞中 ,扩增出长 4 4 5bp的鸡IL- 2 c DNA基因 ,以 p UC119为载... 应用逆转录多聚酶链反应 (RT- PCR)技术 ,参照 Sundick发表的鸡白细胞介素 2 (IL- 2 ) c DNA基因序列 ,利用自行设计合成的 1对引物 ,从 Con A活化的 5周龄 HWL - SPF鸡脾细胞中 ,扩增出长 4 4 5bp的鸡IL- 2 c DNA基因 ,以 p UC119为载体 ,克隆了扩增片段 ,经酶切鉴定及 DNA序列测定 ,确认为鸡 IL- 2基因 ,该序列与 Sundick报道的结果一致。对氨基酸的比较发现 ,鸡 IL - 2与牛 IL - 2和 IL - 15之间分别存在着 2 4 %及4 4 %的同源性 ,该基因系国内首次获得经序列测定证实的鸡 IL- 2 c DNA基因 ,这为今后进一步进行鸡 IL- 展开更多
关键词 鸡IL-2 分子克隆 RT-PCR 免疫传剂 预防接种
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大肠杆菌热敏毒素LT克隆基因的PCR定点诱变和重组表达的构建 被引量:1
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作者 马兴元 郑文云 +2 位作者 赵玉军 姜力 朱平 《中国预防兽医学报》 CAS CSCD 北大核心 2002年第6期427-431,共5页
为使大肠杆菌热敏毒素LT的毒素活性丧失的同时仍保留其较强的免疫原性 ,实验依据LT基因的同源序列设计并合成 5条引物 ,采用突出末端PCR法和重组PCR法 ,将克隆于质粒pEWD2 99上的LT基因 ,分别引入BamHI、Ndel和xhol等位点 ,扩增出带有上... 为使大肠杆菌热敏毒素LT的毒素活性丧失的同时仍保留其较强的免疫原性 ,实验依据LT基因的同源序列设计并合成 5条引物 ,采用突出末端PCR法和重组PCR法 ,将克隆于质粒pEWD2 99上的LT基因 ,分别引入BamHI、Ndel和xhol等位点 ,扩增出带有上述RE位点且含有m7和m112 突变点的约 110 0bp和约 80 0bp的DNA片段和不含突变点的约 30 0bp的DNA片段 ,使控制LT毒力活性中心的第 7位和第 112位氨基酸的碱基分别发生诱变 ,各DNA片段经分离、纯化、RE酶切和DNA连接酶连接后 ,插入经BamHI和Xhol双酶切的线性化的高效表达载体pGEX_4T_1的多克隆位点区 ,使LTm基因置于pTac强启动子下且与Thrombin蛋白基因融合表达 ,将连接产物转化入JM10 5菌株 ,挑出可疑阳性菌落 ,提取质粒 ,经酶切鉴定、PCR鉴定和DNA序列分析 ,证明读框正确 ,序列正确 ,获得了pGEX_4T_1(LTm7和pGEX_4T_1(LTm112 两个重组表达质粒。为运用基因工程手段大量生产人和动物大肠杆菌流行性腹泻的疫苗抗原和幽门螺杆菌疫苗的粘膜免疫佐剂 ,完成了基因水平的工作。 展开更多
关键词 大肠杆菌 热敏毒素 LT克隆基因 PCR定点诱变 重组表达 粘膜免疫传剂 基因工程
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Current use of immunosuppressive agents in inflammatory bowel disease patients in East China 被引量:6
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作者 Li-Juan Huang Qin Zhu +1 位作者 Min Lei Qian Cao 《World Journal of Gastroenterology》 SCIE CAS CSCD 2009年第24期3055-3059,共5页
AIM:To investigate immunosuppressive agents used to treat inflammatory bowel disease(IBD)in East China. METHODS:A retrospective review was conducted, involving 227 patients with IBD admitted to Sir Run Run Shaw Hospit... AIM:To investigate immunosuppressive agents used to treat inflammatory bowel disease(IBD)in East China. METHODS:A retrospective review was conducted, involving 227 patients with IBD admitted to Sir Run Run Shaw Hospital,College of Medicine,Zhejiang University from June 2000 to December 2007.Data regarding demographic,clinical characteristics and immunosuppressants usage were analyzed. RESULTS:A total of 227 eligible patients were evaluated in this study,including 104 patients with Crohn’s disease and 123 with ulcerative colitis.Among the patients,61 had indications for immunosuppressive agents use.However,only 21 (34.4%)received immunosuppressive agents.Among the 21 patients,6(37.5%)received a subtherapeutic dose of azathioprine with no attempt to increase the dosage.Of the 20 patients that received immunosuppressive agent treatment longer than 6 mo,15 patients went into remission,four patients were not affected and one relapsed.Among these 20 patients,four patients suffered from myelotoxicity and one suffered from hepatotoxicity.CONCLUSION:Immunosuppressive agents are used less frequently to treat IBD patients from East China compared with Western countries.Monitoring immunosuppressive agent use is recommended to optimize dispensation of drugs for IBD in China. 展开更多
关键词 Inflammatory bowel disease Immunosuppressive agents AZATHIOPRINE
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Human endogenous retroviruses and cancer 被引量:3
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作者 María Gonzalez-Cao Paola Iduma +3 位作者 Niki Karachaliou Mariacarmela Santarpia Julià Blanco Rafael Rosell 《Cancer Biology & Medicine》 SCIE CAS CSCD 2016年第4期483-488,共6页
Human endogenous retroviruses(HERVs) are retroviruses that infected human genome millions of years ago and have persisted throughout human evolution. About 8% of our genome is composed of HERVs, most of which are nonf... Human endogenous retroviruses(HERVs) are retroviruses that infected human genome millions of years ago and have persisted throughout human evolution. About 8% of our genome is composed of HERVs, most of which are nonfunctional because of epigenetic control or deactivating mutations. However, a correlation between HERVs and human cancer has been described and many tumors, such as melanoma, breast cancer, germ cell tumors, renal cancer or ovarian cancer, express HERV proteins, mainly HERV-K(HML6) and HERV-K(HML2). Although the causative role of HERVs in cancer is controversial, data from animal models demonstrated that endogenous retroviruses are potentially oncogenic. HERV protein expression in human cells generates an immune response by activating innate and adaptive immunities. Some HERV-derived peptides have antigenic properties. For example, HERV-K(HML-6) encodes the HER-K MEL peptide recognized by CD8+ lymphocytes. In addition, HERVs are twoedged immunomodulators. HERVs show immunosuppressive activity. The presence of genomic retroviral elements in host-cell cytosol may activate an interferon type I response. Therefore, targeting HERVs through cellular vaccines or immunomodulatory drugs combined with checkpoint inhibitors is attracting interest because they could be active in human tumors. 展开更多
关键词 HERVs CANCER INTERFERON IMMUNOTHERAPY
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