IDENTIFICATION OF DNA-SEQUENCES REQUIRED FOR TRANSCRIPTION OF THE HUMAN ALPHA-1-GLOBIN GENE IN A NEW SV40 HOST-VECTOR SYSTEM
IDENTIFICATION OF DNA-SEQUENCES REQUIRED FOR TRANSCRIPTION OF THE HUMAN ALPHA-1-GLOBIN GENE IN A NEW SV40 HOST-VECTOR SYSTEM
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DOI：
10.1016/0092-8674(81)90411-6
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发表时间：
1981-01-01
期刊：
影响因子：
64.5
通讯作者：
MANIATIS, T
中科院分区：
文献类型：
--
作者：
MELLON, P;PARKER, V;MANIATIS, T
描述了一种用于研究克隆的真核基因的转录的快速和简单的方法;它涉及用含有SV 40病毒复制起点但缺乏病毒转录所需区域的质粒pBR 322的衍生物（SV-ORI载体）来转染SV 40转化的猴细胞系（COS细胞）。因为COS细胞产生SV 40 T抗原并且允许SV 40病毒复制，所以转染的SV-ORI质粒复制到高拷贝数。携带人α-ORI质粒的SV-ORI质粒珠蛋白基因也在COS细胞中复制。α-珠蛋白基因被忠实地转录以产生高水平的RNA，其被精确地加工以产生真实的α-珠蛋白。珠蛋白mRNA。该转录系统用于证明位于mRNA加帽位点上游55-87个碱基对的序列是α-半乳糖苷酶的有效转录所需的。COS细胞珠蛋白基因。
A rapid and simple method for studying the transcription of cloned eukaryotic genes is described; it involves transfecting SV40-transformed monkey cell lines (COS cells) with derivatives of the plasmid pBR322 that contain the SV40 viral replication origin but lack regions necessary for viral transcription (SV-ORI vectors). Because COS cells produce SV40 T antigen and are permissive for SV40 viral replication, transfected SV-ORI plasmids replicate to a high copy number. SV-ORI plasmids carrying a human .alpha.-globin gene are also replicated in COS cells. The .alpha.-globin gene is faithfully transcribed to produce high levels of RNA, which is accurately processed to produce authentic .alpha.-globin mRNA. This transcription system was used to demonstrate that a sequence located 55-87 base pairs upstream from the mRNA capping site is required for efficient transcription of the .alpha.-globin gene in COS cells.