Difference between revisions of "Part:BBa K1017404"
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[[File:NCTU sRNA-2 structure predict.png|center]] | [[File:NCTU sRNA-2 structure predict.png|center]] | ||
| + | ===Effect on E. Coli Growth=== | ||
| + | To test whether or not our sRNA would effect the growth of E. Coli, we compared the growth of E. Coli with PSB1C3 (without RFP) and the E. Coli growth with Pcons + sRNA. It can be observed from the following figure that the resultant growth curves are similar, showing no signs of growth interference. This result proves that our sRNA regulated system can be integrated into bacteria. | ||
| + | [[File:NCTU_sRNA_growth.JPG|center|600px|The growth curve of the E. Coli with sRNA shows uninterrupted growth curve | ||
| + | that is similar to the growth curve of the E. Coli with PSB1C3.]] | ||
===Quantitative data showing the Part or Device function=== | ===Quantitative data showing the Part or Device function=== | ||
===Acknowedgment of sources and references=== | ===Acknowedgment of sources and references=== | ||
Revision as of 16:00, 29 September 2013
sRNA-2
Description of function
Small RNA is an important role on gene regulation. It will interact with the targeted mRNA by imperfect basepairing, reducing the translational efficiency and recruiting chaperones such as Hfq for translational termination.Since we choose sRNA to be part of our regulated-system, we need to prevent the sRNA from effecting undesired genes, so it is necessarily to design artificial sRNA that targets specifically to the desired genes.
The following figure depicts the mechnism of sRNA. The sRNA basepair with the target mRNA and the ribosome can't bind the mRNA. Therefore, the translation would be inhibited.
How we design
We first picked the sRNA from a library of artificial sRNA that was constructed by fusing a randomized antisense domain of Spot42, the scaffold that is known to recruit the RNA chaperons. As we want the sRNA we picked can specifically complementary to the SD sequence which in our designed RBS(BBa_K1017202[1]), we picked the one which contains a consensus sequence, 5’-CCCUC-3’. Our artificial sRNA also has three stem-loop double stranded RNA structures, and the loop closest to the 3’ terminus is complementary to a sequence preceding the initiation codon of mRNA, so that it can prevent the ribosome from binding to the initiation condon, so the translation would be repressed.
Effect on E. Coli Growth
To test whether or not our sRNA would effect the growth of E. Coli, we compared the growth of E. Coli with PSB1C3 (without RFP) and the E. Coli growth with Pcons + sRNA. It can be observed from the following figure that the resultant growth curves are similar, showing no signs of growth interference. This result proves that our sRNA regulated system can be integrated into bacteria.
Quantitative data showing the Part or Device function
Acknowedgment of sources and references
Torsten Waldminghaus, Nadja Heidrich, Sabine Brantl and Franz Narberhaus .(2012). Engineering Artificial Small RNAs for Conditional Gene Silencing in Escherichia coli ,1: 6–13 Sequence and Features
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