pCryptDel4.8 BioVector?質(zhì)粒 – 基于CRISPR的自我清除型隱型質(zhì)粒消除載體
- 價(jià) 格：￥99850
- 貨 號(hào)：BioVector? pCryptDel4.8
- 產(chǎn) 地：北京
- BioVector NTCC典型培養(yǎng)物保藏中心
- 聯(lián)系人：Dr.Xu, Biovector NTCC Inc.
電話(huà)：400-800-2947 工作微信:1843439339 (QQ同號(hào))
手機(jī)：18901268599
地址：北京
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BioVector? pCryptDel4.8 Plasmid – A CRISPR?Based Self?Curing Vector for Removal of Cryptic Plasmids
BioVector? pCryptDel4.8 質(zhì)粒 – 基于CRISPR的自我清除型隱型質(zhì)粒消除載體
1. Product Description / 產(chǎn)品描述
BioVector? pCryptDel4.8 is a CRISPR?Cas9?based plasmid engineered for the one?step elimination of endogenous cryptic plasmids from Escherichia coli Nissle 1917 (EcN) and related bacterial hosts. It employs a dual?gRNA strategy: one guide RNA targets the cryptic plasmid (e.g., pMUT2), while a second self?targeting gRNA removes the BioVector? pCryptDel4.8 plasmid itself after successful curing. The inclusion of the RelB antitoxin enhances plasmid stability during the curing process.
BioVector? pCryptDel4.8 是一款基于CRISPR?Cas9系統(tǒng)的工程化質(zhì)粒，專(zhuān)用于一步法清除大腸桿菌Nissle 1917（EcN）及相關(guān)宿主中的內(nèi)源隱型質(zhì)粒。該質(zhì)粒采用雙gRNA策略：一條gRNA靶向隱型質(zhì)粒（如pMUT2），另一條自我靶向gRNA在清除完成后消除BioVector? pCryptDel4.8質(zhì)粒本身。同時(shí)攜帶RelB抗毒素，以增強(qiáng)清除過(guò)程中的質(zhì)粒穩(wěn)定性。
2. Product Specifications / 產(chǎn)品規(guī)格
| Parameter / 參數(shù) | Specification / 規(guī)格 |
|---|---|
| Plasmid Name / 質(zhì)粒名稱(chēng) | BioVector? pCryptDel4.8 |
| Vector Backbone / 載體骨架 | pFREE |
| Total Size / 總大小 | 7621 bp |
| Vector Type / 載體類(lèi)型 | Bacterial Expression, CRISPR, Synthetic Biology |
| Bacterial Resistance / 抗性標(biāo)記 | Kanamycin (50?μg/mL) |
| Copy Number / 拷貝數(shù) | High Copy |
| Growth Strain / 宿主菌 | DH5α |
| Growth Temperature / 培養(yǎng)溫度 | 37?℃ |
3. Insert Information / 插入片段信息
3.1 RelB Antitoxin / RelB 抗毒素
| Parameter / 參數(shù) | Specification / 規(guī)格 |
|---|---|
| Gene Name / 基因名稱(chēng) | RelB |
| Species / 來(lái)源 | Escherichia coli Nissle 1917 |
| Insert Size / 插入片段大小 | 142 bp |
| GenBank ID | WP_001554710 |
| Promoter / 啟動(dòng)子 | Native promoter from pMUT2 relB/relE operon |
| Cloning Method / 克隆方法 | Gibson Cloning |
3.2 gRNA targeting cryptic plasmid (e.g., pMUT2) / 靶向隱型質(zhì)粒的gRNA
| Parameter / 參數(shù) | Specification / 規(guī)格 |
|---|---|
| Gene Name / 基因名稱(chēng) | gRNA against target cryptic plasmid |
| Species / 來(lái)源 | Synthetic |
| Insert Size / 插入片段大小 | 30 bp |
| Promoter / 啟動(dòng)子 | Prha (rhamnose?inducible promoter) |
| Cloning Method / 克隆方法 | Restriction Enzyme (BsaI) |
4. Applications / 應(yīng)用
One?step curing of endogenous cryptic plasmids (e.g., pMUT2) from Escherichia coli Nissle 1917 (EcN).
CRISPR?Cas9?based plasmid elimination in bacterial synthetic biology.
Preparation of plasmid?free probiotic strains for engineering and therapeutic applications.
Removal of cryptic plasmids prior to constructing neoantigen delivery vectors.
一步法清除大腸桿菌Nissle 1917（EcN）中的內(nèi)源隱型質(zhì)粒（如pMUT2）。
基于CRISPR?Cas9的細(xì)菌合成生物學(xué)質(zhì)粒清除。
制備無(wú)隱型質(zhì)粒的益生菌菌株，用于工程改造及治療應(yīng)用。
在構(gòu)建新抗原遞送載體前預(yù)先清除隱型質(zhì)粒。
5. Storage and Handling / 儲(chǔ)存與使用建議
Shipping Format / 運(yùn)輸形式: Bacterial stab or purified DNA
Storage / 儲(chǔ)存條件: Store at –20?℃ (purified DNA) or 4?℃ (bacterial stab, short term)
Recommended Strain for Propagation / 推薦擴(kuò)增菌株: DH5α at 37?℃ with kanamycin (50?μg/mL) selection
6. Citation / 引用說(shuō)明
When using BioVector? pCryptDel4.8 in your research, please cite the original publication(s) and include the following acknowledgement in your Materials and Methods:
“BioVector? pCryptDel4.8 was used in this study (obtained from BioVector).”
在研究中引用BioVector? pCryptDel4.8時(shí)，請(qǐng)引用原始文獻(xiàn)，并在材料與方法部分注明：
“本研究使用了BioVector? pCryptDel4.8（由BioVector提供）?！?/p>
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