T7 RNA Polymerase with Genetically Engineered Improved Thermostability

Thermo T7 RNA Polymerase «TT7»


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Thermo T7 RNA Polymerase is a version of wild-type T7 RNA Polymerase, into which a point mutation is introduced through genetic engineering to improve thermostability. Compared to wild-type T7 RNA Polymerase, this polymerase has higher thermostability, allowing reactions even at 50°C It also has improved specific activity at 37°C.

Application

 

  • Preparation of Extensively Labeled RNA Probes
  • Preparation of Precursors for RNA Splicing Reactions
  • Production of Capped mRNA with a Cap Analog as the Primer
  • Preparation of mRNA for In Vitro Translation

1.Comparison of Thermostability
We compared thermostability between native T7 RNA Polymerase and this enzyme. Remaining enzymatic activities were measured after incubation of Native T7 RNA Polymerase at 48°C and Thermo T7 RNA Polymerase at 50°C for each reaction time. Results showed that activity decreased to approximately 1/10 after 5-minute incubation at 48°C for the former, whereas activity had barely decreased even after 15-minute incubation at 50°C with Thermo T7 RNA Polymerase.


Half-life at 50°C
Native T7 RNA Polymerase<1.9min.
Thermo T7 RNA Polymerase 84.5min.

2. In Vitro transcription Reaction at High Temperature
We compared in vitro transcription using Native T7 RNA Polymerase and Thermo T7 RNA Polymerase.
20 U of RNase Inhibitor and 25 U of both enzymes were added using 0.5µg of plasmid containing the T7 promoter sequence as a template to incubate at 37-51°C for 60 minutes, followed by electrophoresis of reaction products. Results showed that Thermo T7 RNA Polymerase allows transcription even at 51°C.

 

Feature and Advantages

Improved Thermostability
High thermostability allows reactions even at 50°C.

Principle

Similar to T3 RNA Polymerase

Product List

Product Name Cat# Quantity Price

Thermo T7 RNA Polymerase ((TT7)) DataSheet ,

TYB-TRL-201

1*7500 UNIT ¥16,100
$215

Thermo T7 RNA Polymerase ((TT7))(Highly concentrated) DataSheet ,

TYB-TRL-252

1*50000 UNIT ¥93,400
$1246

To be used for research only. DO NOT use for human gene therapy or clinical diagnosis.


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