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| | + | A new version of er-(n2)oxStayGold(c4), which gives a higher expression level. The StayGold gene has codon optimization for mammalian expression. For brighter ER imaging if permitted |
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| − | RNA polymerase in ''E. coli'' 两个单引号是斜体. relies on combined with σ factors to recognize promoter sequences. Although σ70 factor is responsible for the transcription of most of the genes in the genome. ''E. coli'' has other six kinds of alternative σ factors that are triggered in stressful conditions. By switching σ factors, bacteria can dramatically change the whole transcription pattern in order to express specific proteins to help themself survive in a bad ''day''.
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| − | Among the six alternative σ factors, σS factor recognizes DNA sequences that are very similar to the typical promoter sequences recognized by σ70. Therefore, it is thought that σS promoters rely on upstream elements<ref>What makes an Escherichia coli promoter sigma(S) dependent? Role of the -13/-14 nucleotide promoter positions and region 2.5 of sigma(S). Becker G, Hengge-Aronis R. Mol Microbiol, 2001 Mar;39(5):1153-65. PMID:11251833</ref> 这里插入全部参考文献，会自动产生列表和链接 , which are protein-binding DNA sequences distributed upstream, to show their dependence on σS factors.
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| − | We noticed that '''there’s no standard nor well-characterized σS promoter in the registry''' 三个单引号是加黑. Therefore, we tested the core regions of several σS promoters and examined where they can be recognized by σS factors independently. Among them, the core region of the promoter controlling the expression of dps in ''E. coli'' became our favorite, shows highest strength as a σS promoter.
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| − | ===Usage and Biology===
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| − | The sequence of this aWASD ewASDSA ewAD fsEEE AsaD product (gp) 5.7 ([https://parts.igem.org/Part:BBa_Z0141 Z0141], [https://parts.igem.org/Part:BBa_K4162050 K4162050]) 方括号是链接 , that acts as an efficient inhibitor of σS-dependent transcription. We image we could develop a circuit simultaneously controls the expression of all the proteins whose transcription is regulated by the expression of gp5.7.
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| − | ===Worth to Notice===
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| − | This part is the simplified form of dps promoter. It’s short and functions as a σS promoter independently. It can be freely fused with other regulatory parts, such as operators, to create parts with novel properties.
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| − | ===Characterization===
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| − | ====Growth Curve and GFP Concentration 要换这个标题 ====
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| − | Plasmids (Figure 1,2) 图无法自动引用 were constructed and respectively transformed into DH5α ''E. coli''. Three independent colonies of each strain were picked out and cultivated at 37℃ overnight in LB culture medium with ampicillin. Then, the bacteria solution was diluted to 1% with 100 ml LB culture medium and shaked for 11.5 hours in 37℃. Samples of bacteria solution were collected every 0.5 hour, and then stored in 4℃ for measurement later using a fluorescent plate reader. After 11.5 hours, all the samples were transfered from eppendorf tubes into wells on 96-well plates. OD600 and fluorescence intensity (excitation: 488 nm, emission: 530 nm) of each sample was measured twice with a plate reader.
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| − | [[File:T--Fudan--K4162001fig1.png|400px|thumb|none|'''Figure 1. GFP controlled by dps promoter (pdps).''' The RBS is part B0034. The GFP is part E0040. The terminator is part BBa_0015.]]
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| − | 两个方括号内的图，分别是为 文件名 宽度 是否缩略图 是否悬浮，可以只改文件名的部分来用，图标题 图注
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| − | As shown in Figure 4, the OD600 (1 OD equals to 10^8 950-nm diameter silica nanoparticles from [https://nanocym.com/ NanoCym]) growth curves of both bacteria share similar trend. Both bacterial strains are at exponential stage before 5 hours in culturre, and then reach a plateau. The overall OD600 curve fro
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| − | ====Compare six σS promoters 一个部分结束，要改 ====
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| − | inal p'''dps'''.
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| − | ====T7 gene product 5.7 inhibition test====
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| − | Gene produ mplex<ref>T7 phage factor required for managing RpoS in Escherichia coli. Tabib-Salazar A, Liu B, Barker D, Burchell L, Qimron U, Matthews SJ, Wigneshweraraj S. Proc Natl Acad Sci U S A, 2018 Jun 5;115(23):E5353-E5362. PMID:29789383</ref>. 又一个参考文献在一对尖括号内，例子里的参考文献格式是pubmed直接copy的
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| − | [[File:T--Fudan--K4162001fig7.png|400px|thumb|none|'''Figure 7. Plasmid for inducible T7 gp5.7 expression.''']]
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| − | We constructed the plasmid shown in Figure 7, T7 gp5.7 regulate help us understand this process https://2021.igem.org/Team:Fudan/Model 这个不加方括号是有文字有链接，加了方括号只有链接。
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| − | 下面的部分小心改part号 最后的参考文献是自动生成的
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A new version of er-(n2)oxStayGold(c4), which gives a higher expression level. The StayGold gene has codon optimization for mammalian expression. For brighter ER imaging if permitted
