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EBV特异性CTL相关TCR Vα-pIRES-TCR Vβ重组质粒的构建及体外表达 被引量:1
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作者 吴秀丽 李扬秋 +5 位作者 朱康儿 杨力建 陈少华 Piotr Grabarczyk grzegorz k.przybylski Christian A.Schmidt 《免疫学杂志》 CAS CSCD 北大核心 2008年第4期412-416,共5页
目的构建EBV特异性细胞毒T细胞(CTL)相关的TCR Vα-pIRES-TCR Vβ真核双表达质粒。方法通过RT-PCR和基因扫描技术分析EBV特异性CTL克隆的T细胞受体(TCR)Vα和Vβ谱系表达情况及T细胞克隆性,确定其优势表达的TCR Vα15和TCR Vβ1亚家族... 目的构建EBV特异性细胞毒T细胞(CTL)相关的TCR Vα-pIRES-TCR Vβ真核双表达质粒。方法通过RT-PCR和基因扫描技术分析EBV特异性CTL克隆的T细胞受体(TCR)Vα和Vβ谱系表达情况及T细胞克隆性,确定其优势表达的TCR Vα15和TCR Vβ1亚家族基因的核苷酸序列(包括CDR3互补决定区),PCR扩增出TCR Vα15和TCR Vβ1基因后,分别定向克隆入真核双表达载体pIRES,构建双顺反子重组质粒TCR Vα-pIRES-TCR Vβ,转基因技术将其转染A549细胞和Molt4细胞,通过RT-PCR、实时定量PCR、流式细胞术和蛋白印迹分析检测TCR Vα15和TCR Vβ1基因的表达。结果酶切分析和核酸序列测定证实重组质粒构建正确,转染重组质粒的细胞能够表达TCR Vα15和TCR Vβ1的mRNA及蛋白。结论成功构建了EBV特异性的TCR Vα-pIRES-TCR Vβ双顺反子真核表达质粒,并在体外实现了共表达,为进一步的实验研究奠定了基础。 展开更多
关键词 EBV 转染 细胞毒T细胞 双顺反子
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Mouse models to study genes involved in hematological malignancies
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作者 grzegorz k.przybylski 《Blood Science》 2020年第2期50-53,共4页
Hematological malignancies,including leukemia and lymphoma,consist a group of highly heterogeneous neoplasms characterized by numerous genetic lesions specific for the type of the disease.In order to understand,the ro... Hematological malignancies,including leukemia and lymphoma,consist a group of highly heterogeneous neoplasms characterized by numerous genetic lesions specific for the type of the disease.In order to understand,the role of a particular alteration in the development of a malignancy functional studies have to be carried out in vitro,in cell lines derived from primary cancer cells.Further efforts to understand the mechanisms underlying blood disorders including malignant transformation and progression relies on model organism research.Numerous transgenic mouse models,carrying human oncogenes have been generated resembling distinct types of hematological disorders.Recent technological advances revolutionized the generation of animal models making it much easier,faster,and precise.The introduction of the CRISPR-Cas9 technology allows for rapid generation of novel knockout or transgenic animals,and the development of conditional site-and time-specific Cre-Lox gene targeting technology,allows studying the function of genes which are relevant to normal hematopoiesis and development of hematological malignancies,but lethal when knocked out in embryonic cells.Besides the studies on gene function,mouse models of human leukemia allow for discovery and testing of novel antileukemic drugs.These new technologies are deepening our understanding of disease pathophysiology and treatment resistance,as well as are leading to novel therapeutic strategies for improved outcomes in patients. 展开更多
关键词 Mouse model CRISPR/Cas9 CRE-LOX LEUKEMIA LYMPHOMA
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