Part:BBa_K283047

T7 RNA polymerase

This part codes for the T7 RNA polymerase.This part does not pose any biological threat.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 1693
  

IPTG inductive effect

In our project,we built recombinant plasmids and transformed them into brewer yeast to produce dsRNA. Because of the lack of T7 RNA promoter, we transformed a plasmid which contained T7 RNA polymerase gene. And IPTG could induce the expression of T7 RNA polymerase gene. We tested the IPTG inductive effect in T7 RNA polymerase. In our project,we built the yeast which contained two parts to produce dsRNA. Meanwhile,IPTG could induce the operation of T7 RNA polymerase.Therefore,we could test the concentration of dsRNA to estimate the inductive effect. After the test of IPTG induction,we firmly believed that we could make use of the best IPTG concentration and induction time lengths to get the minimum cost in agriculture.

The inductive effect of different concentration IPTG

We tested dsRNA concentration with the induction of 0.1,0.2,0.3,0.4,0.5,0.6 mmol/L IPTG after 4 hours

Figure 1.Concentration of dsRdRp differs under different concentration of IPTG. The concentration of dsRdRp continues to increase till the concentration of IPTG reaches to 0.4 mmol/L

The inductive effect of different time length

We tested dsRNA concentration with the induction time length from 0 to 9 hours. And we tested three group which involved concentration.

Figure 2.Concentration of dsRdRp changes with three different concentration of IPTG over time. Induced by IPTG about 4 hours, the concentration of dsRdRp appears to decrease or remain unchanged