Part:BBa_K1960102
sRNA, which can silence chloramphenicol resistance
RNA silencing cassettes, which can silence chloramphenicol resistance.
Usage and Biology
Figure 1. The diagram of synthetized RNA silencing cassettes and control.
We designed and synthetized RNA silencing cassettes and constructed it into pSB1AC3, an official plasmid which contained ampicillin resistance gene and chloramphenicol resistance gene. Also, we used pSB1AC3+mRFP as control. Then we transfer the plasmids into chemical competent cell (DH5α) and add chloramphenicol of 50μg/ml to detect the knockdown effectiveness through the phenotype of survival index. We continuously measured the OD600.
Figure 2. The growth curves of bacteria with the sRNA cassettes or mRFP on the chloramphenicol of 50ng/μL.
The growth of chemical competent cells(DH5α) with pSB1AC3+sRNA is significant repressed by the chloramphenicol (50μg/mL) compared with the control with pSB1AC3+mRFP (The blue line). It indicated the sRNA's highly knockdown effectiveness. From the result, we can found the OD600 of control group shows S-type growth trend. But the experimental group shows a low level all the time. It suggests the growth of the bacteria which contain the sRNA cassettes significantly repressed by the chloramphenicol, which indicates the sRNA's highly knockdown effectiveness.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 153
Illegal XhoI site found at 159 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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