Difference between revisions of "Part:BBa K3384313:Design"
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===Design Notes=== | ===Design Notes=== | ||
| − | Based on the characterization results, we found that although p<em>prm1</em> possesses three PREs, it has a low induced expression level which indicates a good potential for transformation. And the background expression level of this promoter is low which means it can be used to express some proteins that require lower concentrations in the early stage. Based on the literature, we learned the general effect of PREs on promoter strength. For example, an increase in the copy number of PREs can enhance the induced expression level of the promoter. And a PRE which is aligned in the direction of the promoter may be more effective than a PRE which is in the opposite direction of the promoter. p<em>prm1</em> contains 3 × PREs which are in the opposite direction of the promoter. The modified p<em>prm1</em> is referred to as p<em>prm1</em> Pro whose 3 × PREs are in the same direction of the promoter.Fragment1 and Fragment2 were obtained by PCR amplification using | + | Based on the characterization results, we found that although p<em>prm1</em> possesses three PREs, it has a low induced expression level which indicates a good potential for transformation. And the background expression level of this promoter is low which means it can be used to express some proteins that require lower concentrations in the early stage. Based on the literature, we learned the general effect of PREs on promoter strength. For example, an increase in the copy number of PREs can enhance the induced expression level of the promoter. And a PRE which is aligned in the direction of the promoter may be more effective than a PRE which is in the opposite direction of the promoter. p<em>prm1</em> contains 3 × PREs which are in the opposite direction of the promoter. The modified p<em>prm1</em> is referred to as p<em>prm1</em> Pro whose 3 × PREs are in the same direction of the promoter.Fragment1 and Fragment2 were obtained by PCR amplification using p<em>prm1</em>-GFP-CYC1 plasmid as the template. p<em>prm1</em> Pro consists of these two fragments.The sequences of Fragment1 and Fragment2 are shown in Table1. |
Latest revision as of 13:47, 27 October 2020
pprm1 Pro
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
Based on the characterization results, we found that although pprm1 possesses three PREs, it has a low induced expression level which indicates a good potential for transformation. And the background expression level of this promoter is low which means it can be used to express some proteins that require lower concentrations in the early stage. Based on the literature, we learned the general effect of PREs on promoter strength. For example, an increase in the copy number of PREs can enhance the induced expression level of the promoter. And a PRE which is aligned in the direction of the promoter may be more effective than a PRE which is in the opposite direction of the promoter. pprm1 contains 3 × PREs which are in the opposite direction of the promoter. The modified pprm1 is referred to as pprm1 Pro whose 3 × PREs are in the same direction of the promoter.Fragment1 and Fragment2 were obtained by PCR amplification using pprm1-GFP-CYC1 plasmid as the template. pprm1 Pro consists of these two fragments.The sequences of Fragment1 and Fragment2 are shown in Table1.
Source
Saccharomyces cerevisiae
References
[1] Heiman, M. G., and Walter, P. (2000) Prm1p, a pheromone-regulated multispanning membrane protein, facilitates plasma membrane fusion during yeast mating, The Journal of cell biology 151, 719-730.
[2] Sengupta, P., and Cochran, B. H. (1990) The Pre and Pq Box Are Functionally Distinct Yeast Pheromone Response Elements, Molecular and Cellular Biology 10, 6809-6812.