BioVector? pAJM.773細(xì)菌生物傳感器質(zhì)粒 / pAJM.773 Bacterial Biosensor Plasmid

通用定義 / General Definition:

BioVector? pAJM.773 是一種源自麻省理工學(xué)院 (MIT) Voigt 實(shí)驗(yàn)室 Marionette 傳感器集合 的合成生物學(xué)載體。該質(zhì)粒專門用于在非 Marionette 標(biāo)準(zhǔn)大腸桿菌株中構(gòu)建高效的香草酸 (Vanillic acid / Van) 生物傳感器。它是一個(gè)“全合一” (All-in-one) 載體，同時(shí)編碼了轉(zhuǎn)錄阻遏蛋白 VanR-AM 和受其調(diào)控的 PVanCC-YFP 報(bào)告模塊。當(dāng)環(huán)境中存在香草酸時(shí)，VanR 蛋白發(fā)生變構(gòu)并從啟動(dòng)子上解離，從而誘導(dǎo)黃色熒光蛋白 (YFP) 的表達(dá)。該質(zhì)粒是研究小分子感應(yīng)、代謝工程控制以及復(fù)雜合成基因線路的基礎(chǔ)元件。

BioVector? pAJM.773 is a synthetic biology vector from the Marionette Sensor Collection, developed by the Voigt Lab at MIT. This plasmid is specifically designed to function as a high-performance Vanillic acid (Van) biosensor in standard (non-Marionette) E. coli strains. It is an "all-in-one" vector, encoding both the transcriptional repressor VanR-AM and the corresponding PVanCC-YFP reporter module. In the presence of vanillic acid, the VanR protein undergoes a conformational change and dissociates from the promoter, thereby inducing the expression of Yellow Fluorescent Protein (YFP). This plasmid is a fundamental component for small-molecule sensing, metabolic engineering control, and complex synthetic genetic circuit design.

BioVector? pAJM.773 技術(shù)說明書 (Plasmid Datasheet)

中文版說明書

1. 產(chǎn)品基本信息

• 產(chǎn)品名稱： BioVector? pAJM.773 傳感器質(zhì)粒

• Addgene 編號(hào)： 108527

  
  

• 感應(yīng)誘導(dǎo)劑： 香草酸 (Vanillic acid, Van)

  
  

• 報(bào)告基因： EYFP (增強(qiáng)型黃色熒光蛋白)

  
  

• 抗性基因： 卡那霉素 (Kanamycin)

  
  

• 復(fù)制子： p15A (低拷貝，約 10-15 copies/cell)

2. 培養(yǎng)與擴(kuò)增條件

• 宿主菌： BioVector? NEB Stable 或 DH5-alpha。

• 培養(yǎng)基： BioVector? LB 培養(yǎng)基。

• 抗生素： 50 ug/mL 卡那霉素。

  
  

• 環(huán)境： 37 攝氏度，恒溫振蕩。

3. 載體應(yīng)用

• 小分子檢測(cè)： 在大腸桿菌中實(shí)時(shí)定量監(jiān)測(cè)香草酸的濃度。

• 動(dòng)態(tài)代謝控制： 利用環(huán)境中的代謝產(chǎn)物作為開關(guān)，自動(dòng)調(diào)節(jié)下游代謝途徑的基因表達(dá)。

• 生物邏輯門： 作為 Marionette 系統(tǒng)的 12 個(gè)正交傳感器之一，用于構(gòu)建多輸入邏輯線路。

4. 注意事項(xiàng)

• 宿主要求： 即使在普通的非 Marionette 菌株中也能工作，因?yàn)樗詭Я俗瓒舻鞍妆磉_(dá)框。

• 低拷貝優(yōu)勢(shì)： 采用 p15A 復(fù)制子是為了降低漏表達(dá) (Leaky expression) 并減輕細(xì)胞代謝負(fù)擔(dān)。

English Datasheet

1. General Product Information

• Product Name: BioVector? pAJM.773 Sensor Plasmid

• Addgene ID: 108527

  
  

• Inducer: Vanillic acid (Van)

• Reporter Gene: EYFP (Enhanced Yellow Fluorescent Protein)

  
  

• Bacterial Resistance: Kanamycin (50 ug/mL)

  
  

• Replicon: p15A (Low-copy, approx. 10-15 copies/cell)

2. Culture and Amplification Conditions

• Host Strains: BioVector? NEB Stable or DH5-alpha.

• Culture Medium: BioVector? LB Medium.

• Incubation: 37 degrees Celsius with constant shaking.

3. Applications

• Small-Molecule Detection: Real-time quantitative monitoring of vanillic acid concentration in E. coli.

• Dynamic Metabolic Control: Using metabolic intermediates as triggers to automatically regulate gene expression in downstream pathways.

• Genetic Logic Gates: Serving as one of the 12 orthogonal sensors in the Marionette system for constructing multi-input genetic circuits.

  
  

4. Key Usage Notes

• Strain Compatibility: This vector is functional in standard E. coli strains because it contains its own constitutively expressed repressor (VanR).

• Low Leakage: The use of the p15A origin helps minimize basal (leaky) expression and reduces the metabolic burden on the host cell.

Note: For optimal sensor performance with low background noise, BioVector? recommends using M9 minimal media supplemented with necessary carbon sources. Always verify the induction range and sensitivity of pAJM.773 in your specific host strain using BioVector? high-purity Vanillic acid inducer solutions.

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