Product Name / 產(chǎn)品名稱:
BioVector? pEAQ-HT-DEST1 Plasmid植物雙元表達載體，專為在植物中進行重組蛋白的高水平瞬時表達而設(shè)計
BioVector? pEAQ-HT-DEST1質(zhì)粒

Description / 描述:
The BioVector? pEAQ-HT-DEST1 plasmid is a binary plant expression vector designed for high-level transient expression of recombinant proteins in plants -1. It combines virus-derived sequences to enhance translation efficiency with a Gateway? recombination cassette for facile cloning -1-4. Developed from the pEAQ (Easy and Quick) vector series, this plasmid enables rapid production of recombinant proteins at yields up to milligrams per gram of fresh leaf mass upon agroinfiltration -1. The vector is widely used for molecular pharming, protein subcellular localization studies, and functional characterization of plant and pathogen proteins -1-8.
BioVector? pEAQ-HT-DEST1 質(zhì)粒是一種植物雙元表達載體，專為在植物中進行重組蛋白的高水平瞬時表達而設(shè)計 -1。它結(jié)合了病毒來源的序列以增強翻譯效率，并包含Gateway?重組盒以便于克隆 -1-4。該質(zhì)粒源自pEAQ載體系列，通過農(nóng)桿菌浸潤法可在植物中實現(xiàn)高達毫克級/克鮮重的重組蛋白產(chǎn)量 -1。該載體廣泛應(yīng)用于分子農(nóng)業(yè)、蛋白質(zhì)亞細胞定位研究以及植物和病原體蛋白的功能鑒定 -1-8。

Original Source / 原始來源:
Developed by the Lomonossoff Lab, John Innes Centre, UK / 由英國約翰英納斯中心Lomonossoff實驗室開發(fā)

Key Publications / 關(guān)鍵文獻:
Sainsbury F, et al. pEAQ: versatile expression vectors for easy and quick transient expression of heterologous proteins in plants. Plant Biotechnol J. 2009;7(7):682-693.

Peyret H, Lomonossoff GP. The pEAQ vector series: the easy and quick way to produce recombinant proteins in plants. Plant Mol Biol. 2013;83(1-2):51-58.

Plasmid Type / 質(zhì)粒類型:

• Binary plant expression vector / 植物雙元表達載體

• Gateway? destination vector / Gateway?目的載體

• Transient expression vector / 瞬時表達載體

Species / 物種:

• Agrobacterium tumefaciens (for plant transformation) / 根癌農(nóng)桿菌（用于植物轉(zhuǎn)化）

• Escherichia coli (for propagation) / 大腸桿菌（用于擴增）

• Plant hosts (especially Nicotiana benthamiana) / 植物宿主（特別是本氏煙）

Vector Backbone / 載體骨架:
pEAQ-HT-derived / pEAQ-HT衍生載體

Total Vector Size / 總載體大小:
~10.2 kb (10,247 bp) -2 / 約10.2 kb

Key Components / 關(guān)鍵組件:

Gene/Insert Details / 基因/插入片段詳情:

Growth Conditions in Bacteria / 細菌生長條件:

Applications / 應(yīng)用:

• High-level Transient Expression / 高水平瞬時表達: Produces recombinant proteins at yields up to mg/g fresh weight in N. benthamiana-1

• Molecular Pharming / 分子農(nóng)業(yè): Production of therapeutic proteins, vaccines, and industrial enzymes -1

• Protein Subcellular Localization / 蛋白質(zhì)亞細胞定位: Used as backbone for fluorescent fusion protein vectors -1-4

• Effector Biology / 效應(yīng)子生物學: Expression of pathogen effectors to study plant-pathogen interactions -8

• Metabolic Engineering / 代謝工程: Reconstruction of biosynthetic pathways in plants -2-6

• Virus-like Particle Production / 病毒樣顆粒生產(chǎn): Production of vaccine candidates and VLPs

Derived Vector Series / 衍生載體系列:
A collection of 15 pEAQ-HT-DEST1-based vectors has been developed for protein subcellular localization studies -1-4:

Quality Control / 質(zhì)量控制:
Plasmid is verified by restriction digestion and sequencing. Full sequence information is available from public databases (NCBI accession: 257196397) -2.
質(zhì)粒經(jīng)過限制性酶切和測序驗證。完整序列信息可從公共數(shù)據(jù)庫獲取。

Storage / 存儲:

• Plasmid is sent as a bacterial stab in agar or as purified DNA. Upon receipt, streak onto a plate with BioVector? Kanamycin (50 μg/mL), grow a single colony, and prepare a glycerol stock for long-term storage at -80°C.

• 質(zhì)粒以細菌穿刺菌或純化DNA形式提供。收到后，應(yīng)在含 BioVector? 卡那霉素 (50 μg/mL) 的平板上劃線，挑取單克隆，并制備甘油菌于-80°C長期保存。

Plasmid Reconstitution Protocol / 質(zhì)粒溶解復(fù)配方案:

1. Centrifuge at 5,000×g for 5 min. / 5,000×g離心5分鐘。

2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA. / 小心打開管蓋，加入20 μl無菌水溶解DNA。

3. Close the tube and incubate for 10 minutes at room temperature. / 蓋緊管蓋，室溫孵育10分鐘。

4. Briefly vortex and spin down. / 短暫渦旋并離心收集。

5. Store the plasmid at -20°C. / 于-20°C保存質(zhì)粒。

6. Important Note / 重要提示: The intact pEAQ-HT-DEST1 plasmid contains the ccdB gene and must be propagated in E. coli strains resistant to ccdB effects (e.g., DB3.1). For expression clones, use standard cloning strains after Gateway? recombination. / 完整的pEAQ-HT-DEST1質(zhì)粒含有ccdB基因，必須在抗ccdB效應(yīng)的大腸桿菌菌株（如DB3.1）中擴增。對于表達克隆，在Gateway?重組后使用標準克隆菌株。

Citation / 引用:
When using this plasmid, please cite the following publications:
使用此質(zhì)粒時，請引用以下文獻：

Sainsbury F, Thuenemann EC, Lomonossoff GP. pEAQ: versatile expression vectors for easy and quick transient expression of heterologous proteins in plants. Plant Biotechnol J. 2009;7(7):682-693.

For derived fluorescent protein vectors, also cite:
Berthold F, et al. Inside or outside? A new collection of Gateway vectors allowing plant protein subcellular localization or over-expression. Plasmid. 2019;105:102436.

Important Note / 重要提示:

• The pEAQ-HT-DEST1 plasmid is designed for Agrobacterium-mediated transient expression in plants, particularly Nicotiana benthamiana-1-8

• pEAQ-HT-DEST1質(zhì)粒專為農(nóng)桿菌介導(dǎo)的植物瞬時表達設(shè)計，特別適用于本氏煙 -1-8

• The vector contains virus-derived sequences (CPMV UTRs) that dramatically enhance translation efficiency without viral replication -1

• 該載體含有病毒來源的序列，可在無病毒復(fù)制的情況下顯著增強翻譯效率 -1

• The P19 silencing suppressor is encoded within the T-DNA to prevent post-transcriptional gene silencing and maximize expression -1-2

• P19沉默抑制子編碼在T-DNA內(nèi)，可防止轉(zhuǎn)錄后基因沉默，最大化表達水平 -1-2

• The Gateway? cassette enables rapid, directional cloning of genes of interest without restriction enzymes -1-4

• Gateway?盒可實現(xiàn)目的基因的快速、定向克隆，無需限制性內(nèi)切酶 -1-4

• This vector has been successfully used in multiple plant species including pea, lentil, and faba bean, though efficiency varies -8

• 該載體已成功用于多種植物物種，包括豌豆、扁豆和蠶豆，但效率因物種而異 -8

BioVector? Kanamycin
BioVector? 卡那霉素

For bacterial selection of the pEAQ-HT-DEST1 plasmid and its derivatives.
用于pEAQ-HT-DEST1質(zhì)粒及其衍生質(zhì)粒的細菌篩選。

BioVector NTCC質(zhì)粒載體菌株細胞蛋白抗體基因保藏中心

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