Use of retroviral vectors for mapping of splice sites in cottontail rabbit papillomavirus.
Use of retroviral vectors for mapping of splice sites in cottontail rabbit papillomavirus.
复制标题
使用逆转录病毒载体绘制棉尾兔乳头瘤病毒剪接位点图谱。
DOI：
10.1099/0022-1317-69-12-3093
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发表时间：
1988
期刊：
影响因子：
--
通讯作者：
R. Mulligan
中科院分区：
文献类型：
--
作者：
N. Dostatni;M. Yaniv;O. Danos;R. Mulligan
将编码病毒早期功能的棉尾兔乳头瘤病毒（CRPV）基因组序列导入逆转录病毒载体，制备病毒早期区cdna。将E6开放阅读框前两个不同长度的对照序列转染到Psi-2细胞中，用释放的逆转录病毒原液感染NIH3T3细胞。从g418耐药病毒感染细胞与Cos细胞融合后提取原病毒序列，在大肠杆菌中扩增为质粒并进行分析。核苷酸测序显示，仅包含早期编码区的构建体中使用的剪接信号与表达crpv的肿瘤中相同，将E1的开头连接到E2的中间。另一方面，在包含大部分长控制区的构建体中，位于该区域5'端7810位置的剪接供体位点被非常有效地利用，完全排除了1371位置的供体位点的使用。含有从Moloney小鼠白血病病毒启动子转录的CRPV序列的构建体在DNA转染后不能转化小鼠成纤维细胞。
Cottontail rabbit papillomavirus (CRPV) genomic sequences coding for virus early functions were introduced into a retroviral vector in order to produce cDNAs of the viral early region. Two constructs differing in the length of control sequences preceding the E6 open reading frame were transfected into Psi-2 cells and the released retroviral stock was used to infect NIH3T3 cells. The proviral sequences were rescued from antibiotic G418-resistant virus-infected cells after fusion with Cos cells, amplified as plasmids in Escherichia coli and analysed. Nucleotide sequencing showed that the splicing signals used in the construct containing only the early coding region are the same as in CRPV-expressing tumours, linking the beginning of E1 to the middle of E2. On the other hand, in a construct including most of the long control region a splice donor site located in the 5' end of this region, at position 7810, was very efficiently used, totally excluding the use of the donor site at position 1371. None of the constructs containing CRPV sequences transcribed from Moloney murine leukaemia virus promoter was able to transform mouse fibroblasts after DNA transfection.
乳头状瘤和癌中主要棉尾兔乳头瘤病毒晚期 RNA 帽位点的鉴定以及 E2 和次要 E6 编码 mRNA 的定位和定量。
影响因子：
3.7
作者：
Wettstein,FO;Barbosa,MS;Nasseri,M
通讯作者：
Nasseri,M
DOI：
10.1073/pnas.84.8.2150
发表时间：
1987-04-01
影响因子：
11.1
作者：
KORMAN, AJ;FRANTZ, JD;MULLIGAN, RC
通讯作者：
MULLIGAN, RC