Difference between revisions of "Template:Jilin China/Measurement C.js"

Revision as of 00:05, 22 October 2019

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var content_alpha = { prospect:{ title:"Background" },

part: [{ title: "Design", para: [

{ type: "word", cont: "Quantitative characterization of promoter intensity is important and fundamental in synthetic biology. The current common method is to express fluorescent protein downstream of the promoter and measure the promoter's intensity by fluorescence intensity. However, fluorescence intensity often fluctuates greatly with different experimental conditions and Protein sequences. If the promoter is only compared with one characterized promoter, the results may not be convincing enough. Therefore, or we provide a new characterization method with several standard promoters with stable relative expression relationship as a dependable control group.", class: "np5" }, { type: "word", cont: "In this way, the deviation caused by the fluctuation of individual promoter intensity can be reduced to obtain the relative intensity of uncharacterized promoters for a series of promoters, and guide a team to select appropriate promoters to adjust the expression intensity of downstream genes.", class: "np5" },

{ type: "word", cont: "We also built a universal measurement device specifically for the promoter intensity measurement, which allows multiple promoters to be quickly assembled with the same RBS and sfGFP through Golden Gate, respectively.", class: "np5" },

]

}, { title: "Measurement", para: [

{ type: "word", cont: "This year, we used the promoter P(sub)59(subed) that has never been characterized in iGEM, managing to acquire the relative intensity of P(sub)59(subed) to Anderson promoter family, and through which we could regulate the expression intensity of downstream genes by changing promoters.", class: "np5" }, { type: "word", cont: "Therefore, we constructed the promoter intensity measurement device BBa_K3078100, and inserted BBa_J23119, BBa_J23104, BBa_J23108, BBa_J23105 and BBa_J23114 of Anderson promoter family into this part by using Golden Gate assembly, to construct five parts with the same RBS and fluorescent protein (sfGFP). The fluorescence intensity at 528 nm under 485 nm wavelength excitation light was measured. We defined the promoter intensity as Fluorescence/ OD600 (Figure 1). The promoter P(sub)59(subed) has also been constructed into BBa_K3078100 using this method.", class: "np5" },

{ type: "pic", maxClass:"max6", cont: [{ num: 1, adress: "", pre: 100, }

] }, { type: "word", cont: "Figure 1. The fluorescence change (Fluorescence/ OD600) of promoters with time. Different promoters were constructed into the same measurement device, cultured overnight, diluted to OD600=0.02 and then measured the fluorescence at 528 nm with 485nm excitation wavelength.", class: "np6" },

{ type: "word", cont: "According to the data measured by Prof. J. Christopher Anderson. (http://parts.igem.org/Promoters/Catalog/Anderson), the expression intensity of J23114, J23105, J23104, J23108 and J23119 are 0.14, 0.24, 0.51, 0.72, 1, which approximately conformed to a gradient relationship. If the fluorescence intensity were made into an image, we would get a straight line which can be used as the standard of comparison of fluorescence measurement.", class: "np5" },

{ type: "pic", maxClass:"max6", cont: [{ num: 1, adress: "", pre: 100, }

] }, { type: "word", cont: "Figure 2. Orange (0h), red (2h), yellow (4h), dark blue (6h), and green (8h) curves respectively represent the relative intensity of the promoters to BBa_J23119 at the same time, measured by Jilin_China. The green one represents the relative intensity measured by Anderson.", class: "np6" }, ]

}, { title: "Conclusion", para: [

{ type: "word", cont: "However, according to our experiment results, although there were approximate linear relationship among the weak Anderson promoters（BBa_J23108, BBa_J23105 and BBa_J23114）, the strong promoter often unmatched this relationship. This result may be attributed to the small number of reference promoters we selected and the great influence of individual sample fluctuations having on the whole trend. If we selected more promoters to compared with, even if the individual promoter fluctuate, the relative intensity relationship of whole group won’t have huge deviation. Therefore，this kind of measurement will be more meaningful and applicable where there are more reference promoters being measured.", class: "np5" },

]

} ,

]

}