An improved retroviral vector for assaying promoter activity. Analysis of promoter interference in pIP211 vector.
We recently developed a novel promoter assay system using a retroviral vector (pIP200 series). Transcription from the internal promoter, which had been inserted for the promoter assay, was shown to be interfered with by transcription from the upstream long terminal repeat (LTR). Here we report a new high-titer 'self-inactivating' vector, in which transcription interference was virtually eliminated. This new vector was constructed by introducing only a very minor mutation into the 'TATA box' in the 3'-LTR. This mutation was successfully transferred to the 5'-LTR after reverse transcription, yielding a provirus incapable of transcribing viral RNA. The viral titer was not reduced by the mutation, permitting general application of this virus.
Nakajima
K
K
Institute for Protein Research, Osaka University, Japan.
Ikenaka
K
K
Nakahira
K
K
Morita
N
N
Mikoshiba
K
K
D016475:3T3 Cells; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods; D015870:Gene Expression; D005822:Genetic Vectors; D051379:Mice; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000235:genetics; D012190:Retroviridae / Q000235:genetics*
D009838:Oligodeoxyribonucleotides; D012333:RNA, Messenger