Tests of a model of specific contacts in T7 RNA polymerase-promoter interactions

Biochemistry ◽  
1995 ◽  
Vol 34 (2) ◽  
pp. 666-672 ◽  
Author(s):  
Charlie Schick ◽  
Craig T. Martin
Keyword(s):  
Rna Polymerase ◽  
T7 Rna Polymerase ◽  
Rna Polymerase Promoter

In vivo and in vitro activities of point mutants of the bacteriophage T7 RNA polymerase promoter

Biochemistry ◽  
1992 ◽  
Vol 31 (37) ◽  
pp. 9073-9080 ◽  
Author(s):  
Richard A. Ikeda ◽  
G. Sakuntala Warshamana ◽  
Lisa L. Chang
Keyword(s):  
Rna Polymerase ◽  
T7 Rna Polymerase ◽  
Bacteriophage T7 ◽  
Rna Polymerase Promoter

scholarly journals Induction of annexin-1 at transcriptional and post-transcriptional level in rat brain by methylprednisolone and the 21-aminosteroid U74389F

1996 ◽  
Vol 5 (5) ◽  
pp. 370-378 ◽  
Author(s):  
P. H. Voermans ◽  
K. G. Go ◽  
G. J. Ter Horst ◽  
M. H. J. Ruiters ◽  
E. Solito ◽  
...  
Keyword(s):  
Rna Polymerase ◽  
T7 Rna Polymerase ◽  
Appreciable Amount ◽  
Transcriptional Level ◽  
Mrna Transcription ◽  
Annexin 1 ◽  
Vascular Walls ◽  
Adjacent Structures ◽  
Antibody Conjugate ◽  
Rna Polymerase Promoter

Brain tissue of rats pretreated with methylprednisolone or with the 21-aminosteroid U74389F, and that of untreated control rats, was assessed for the expression of annexin-1 (Anx-1) and the transcription of its mRNA. For this purpose Anx-1 cDNA was amplified and simultaneously a T7-RNA-polymerase promoter was incorporated into the cDNA using a polymerase chain reaction (PCR). Then digoxigenin-11-UTP was incorporated into the transcribed cRNA with T7-RNA-polymerase. With this probein situhybridization was carried out on sections of the brain. The probe was visualized by an immunoassay using an antidigoxigenin antibody conjugate. Anx-1 protein was assessed by means of immunohistochemistry using a polyclonal antibody. The various brain areas of the control animals showed an appreciable amount of Anx-1 at mRNA or protein level; on the other hand, the animals which had been pretreated with either steroid, showed a more intense Anx-1 mRNA signal than the controls in many areas. In the pretreated animals Anx-1 immunostaining was unchanged in cortex, basal ganglia, amygdala and septum, but more intense in hippocampus, hypothalamus and thalamus. In ependyma, choroid plexus, meninges, and vascular walls there was no Anx-1 mRNA transcription detectable. An opposite profile was shown by the Anx-1 immunoreactivity, the protein was present in control animals as well as the steroid-pretreated animals, suggesting that here the protein was either from systemic origin, or has diffused from adjacent structures. The results indicated that Anx-1 mRNA transcription is upregulated by either steroid, and that in the untreated animals there is a resting level of Anx-1 mRNA transcription, presumably reflecting physiological influences on Anx-1 expression.

scholarly journals T7Max transcription system

2021 ◽  
Author(s):  
Christopher Deich ◽  
Brock Cash ◽  
Wakana Sato ◽  
Judee Sharon ◽  
Lauren Aufdembrink ◽  
...  
Keyword(s):  
Protein Expression ◽  
Rna Polymerase ◽  
Expression System ◽  
T7 Rna Polymerase ◽  
Dna Templates ◽  
Transcription System ◽  
In Vitro Systems ◽  
In Vitro Protein Expression ◽  
Rna Polymerase Promoter

Efficient cell-free protein expression from linear DNA templates has remained a challenge primarily due to template degradation. Here we present a modified T7 RNA polymerase promoter that acts to significantly increase the yields of both transcription and translation within in vitro systems. The modified promoter, termed T7Max, recruits standard T7 RNA polymerase, so no protein engineering is needed to take advantage of this method. This technique could be used with any T7 RNA polymerase- based in vitro protein expression system. Unlike other methods of limiting linear template degradation, the T7Max promoter increases transcript concentration in a T7 transcription reaction, providing more mRNA for translation.

scholarly journals A reverse-genetics system for Influenza A virus using T7 RNA polymerase

2007 ◽  
Vol 88 (4) ◽  
pp. 1281-1287 ◽  
Author(s):  
Emmie de Wit ◽  
Monique I. J. Spronken ◽  
Gaby Vervaet ◽  
Guus F. Rimmelzwaan ◽  
Albert D. M. E. Osterhaus ◽  
...  
Keyword(s):  
Influenza Virus ◽  
Rna Polymerase ◽  
Influenza A Virus ◽  
Influenza A ◽  
Reverse Genetics ◽  
T7 Rna Polymerase ◽  
Influenza Virus A ◽  
Sense Orientation ◽  
Polymerase I ◽  
Rna Polymerase Promoter

The currently available reverse-genetics systems for Influenza A virus are all based on transcription of genomic RNA by RNA polymerase I, but the species specificity of this polymerase is a disadvantage. A reverse-genetics vector containing a T7 RNA polymerase promoter, hepatitis delta virus ribozyme sequence and T7 RNA polymerase terminator sequence has been developed. To achieve optimal expression in minigenome assays, it was determined that viral RNA should be inserted in this vector in the negative-sense orientation with two additional G residues downstream of the T7 RNA polymerase promoter. It was also shown that expression of the minigenome was more efficient when a T7 RNA polymerase with a nuclear-localization signal was used. By using this reverse-genetics system, recombinant influenza virus A/PR/8/34 was produced more efficiently than by using a similar polymerase I-based reverse-genetics system. Furthermore, influenza virus A/NL/219/03 could be rescued from 293T, MDCK and QT6 cells. Thus, a reverse-genetics system for the rescue of Influenza A virus has been developed, which will be useful for fundamental research and vaccine seed strain production in a variety of cell lines.

Evidence for DNA bending at the T7 RNA polymerase promoter

2000 ◽  
Vol 295 (5) ◽  
pp. 1173-1184 ◽  
Author(s):  
Andrea Újvári ◽  
Craig T Martin
Keyword(s):  
Rna Polymerase ◽  
Dna Bending ◽  
T7 Rna Polymerase ◽  
Rna Polymerase Promoter
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