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

Latest revision as of 20:22, 21 October 2019

/**

* 
* @authors lhx 
* @date    2019-05-30 16:25:38
* @version 1.1
*/

var content_alpha = { prospect:{ title:"Improvement" }, reference:[

"[1] Ikeda R A, Warshamana G S, Chang L L. In vivo and in vitro activities of point mutants of the bacteriophage T7 RNA polymerase promoter[J]. Biochemistry, 1992, 31(37): 9073-80.", "[2] Paul S, Stang A, Lennartz K, et al. Selection of a T7 promoter mutant with enhanced in vitro activity by a novel multi-copy bead display approach for in vitro evolution[J]. Nucleic Acids Res, 2013, 41(1): e29.", "[3] Nie Z, Luo H, Li J, et al. High-Throughput Screening of T7 Promoter Mutants for Soluble Expression of Cephalosporin C Acylase in E. coli[J]. Appl Biochem Biotechnol, 2019.", ], part: [{ title: "Design", para: [

{ type: "word", cont: "T7 promoter is one of the most common promoters. But the intensity of the wild type T7 promoter often can’t meet our demand, so we mutated it and obtained an improved T7 promoter. According to previous studies, we selected the mutation site and constructed three mutated promoters（BBa_K3078012, BBa_K3078013, BBa_K3078014）based on the wild-type T7 promoter (BBa_K2150031) into the measurement vector (BBa_K3078100).", class: "np5" }, { type: "pic", maxClass:"max9", cont: [{ num: 1, adress: "", pre: 100, }

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

] },

]},

{ title: "Measurement", para: [

{ type: "word", cont: "The promoter activity of wild-type and mutant T7 were measured through fluorescence intensity and normalized by OD(sub)600(subed). BBa_K2150031 and the mutated T7 promoter lack of T7 RNA polymerase, so we added IPTG to induce the expression of T7 RNA polymerase downstream of promoter lacUV5 of on the BL21(DE3) genome. As shown in Figure 2, compared to wild type T7, the Mu-T7 (BBa_K30178012) showed the higher intensity.", class: "np5" }, { type: "pic", maxClass:"max6", cont: [{ num: 1, adress: "", pre: 100, }

] },

{ type: "word", cont: "Figure 2. The plasmid was transferred to (ita)E. coli(itaed) BL21, cultured overnight, and diluted OD(sub)600(subed) 0.02. IPTG was added at OD(sub)600(subed) 0.3 to make the final concentration of IPTG reach 1mM to measure at the indicated time.", class: "np5" }

]

},

{ title: "Conclusion", para: [

{ type: "word", cont: "Above all, we constructed wild-type and mutant T7 promoters into the measurement vector (BBa_K3078100) by Golden Gate method, and confirmed by the fluorescence measurement of the expression intensity of BBa_K3078012 was 30% higher than that of wild-type T7 promoters. Additionally, the mutation site didn't form the new recognition site of endonucleases or other enzymes, which may affect the digestion and connection or the other assembly ways.", class: "np5" }

]

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

{ type: "word", cont: "[1] Ikeda R A, Warshamana G S, Chang L L. In vivo and in vitro activities of point mutants of the bacteriophage T7 RNA polymerase promoter[J]. Biochemistry, 1992, 31(37): 9073-80.", class: "np5" },

{ type: "word", cont: "[2] Paul S, Stang A, Lennartz K, et al. Selection of a T7 promoter mutant with enhanced in vitro activity by a novel multi-copy bead display approach for in vitro evolution[J]. Nucleic Acids Res, 2013, 41(1): e29.", class: "np5" }, { type: "word", cont: "[3] Nie Z, Luo H, Li J, et al. High-Throughput Screening of T7 Promoter Mutants for Soluble Expression of Cephalosporin C Acylase in E. coli[J]. Appl Biochem Biotechnol, 2019.", class: "np5" }, ]

}

]

}