Use of second-site homologous recombination to demonstrate that Epstein-Barr virus nuclear protein 3B is not important for lymphocyte infection or growth transformation in vitro
Use of second-site homologous recombination to demonstrate that Epstein-Barr virus nuclear protein 3B is not important for lymphocyte infection or growth transformation in vitro
复制标题
利用第二位点同源重组证明Epstein-Barr病毒核蛋白3B对于体外淋巴细胞感染或生长转化并不重要
DOI：
10.1128/jvi.66.5.2893-2903.1992
复制
发表时间：
1992
影响因子：
5.4
通讯作者：
E. Kieff
中科院分区：
文献类型：
--
作者：
B. Tomkinson;E. Kieff
采用二位点同源重组的方法，制备了将终止密码子插入核蛋白3B （EBNA 3B）开放阅读框的重组eb病毒。这些突变病毒感染和生长转化原代B淋巴细胞，导致淋巴母细胞样细胞系（LCLs）的建立。聚合酶链反应分析和与感染细胞DNA的Southern杂交表明存在突变型EBNA 3B，而不存在野生型EBNA 3B。用亲和纯化的EBNA 3B抗体对lcl进行免疫印迹分析，证实不存在EBNA 3B交叉反应蛋白。病毒从两个被感染的lcl中被重新激活，并依次通过初级B淋巴细胞。新感染的细胞只含有突变的重组病毒。在潜伏（EBNA 3B除外）或裂解周期感染的细胞病毒蛋白表达或潜伏感染转化细胞系的生长方面，平行衍生的突变型重组体和野生型重组体没有发现差异。这些数据表明EBNA 3B蛋白在体外对原发性b淋巴细胞感染、生长转化或裂解病毒感染并不重要。
Recombinant Epstein-Barr viruses with a stop codon inserted into the nuclear protein 3B (EBNA 3B) open reading frame were generated by second-site homologous recombination. These mutant viruses infected and growth transformed primary B lymphocytes, resulting in the establishment of lymphoblastoid cell lines (LCLs). Polymerase chain reaction analysis and Southern hybridizations with infected cell DNA demonstrated the presence of the mutant EBNA 3B and the absence of wild-type EBNA 3B. Immunoblot analysis of the LCLs with affinity-purified EBNA 3B antibodies confirmed the absence of EBNA 3B cross-reactive protein. Virus was reactivated from two of these infected LCLs and serially passaged through primary B lymphocytes. The newly infected cells contained only the mutant recombinant virus. No difference was noted between mutant and wild-type recombinants, derived in parallel, in latent (other than EBNA 3B) or lytic cycle-infected cell virus protein expression or in the growth of the latently infected transformed cell lines. These data indicate that the EBNA 3B protein is not critical for primary B-lymphocyte infection, growth transformation, or lytic virus infection in vitro.
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DOI：
10.1073/pnas.81.23.7632
发表时间：
1984-01-01
期刊：
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES
影响因子：
--
作者：
DAMBAUGH, T;HENNESSY, K;KIEFF, E
通讯作者：
KIEFF, E
DOI：
10.1073/pnas.88.4.1546
发表时间：
1991-02
影响因子：
11.1
作者：
S. Swaminathan;B. Tomkinson;E. Kieff
通讯作者：
S. Swaminathan;B. Tomkinson;E. Kieff
使用重组痘苗病毒证明了针对 Epstein-Barr 病毒核抗原的人类细胞毒性 T 细胞反应。
DOI：
10.1073/pnas.87.8.2906
发表时间：
1990
影响因子：
11.1
作者：
Murray,RJ;Kurilla,MG;Griffin,HM;Brooks,JM;Mackett,M;Arrand,JR;Rowe,M;Burrows,SR;Moss,DJ;Kieff,E
通讯作者：
Kieff,E
影响因子：
3.7
作者：
PETTI, L;SAMPLE, C;KIEFF, E
通讯作者：
KIEFF, E
影响因子：
5.3
作者：
KOZAK, M
通讯作者：
KOZAK, M