BioVector? pSRK-GM 廣宿主誘導(dǎo)型表達(dá)載體 / BioVector? pSRK-GM Broad-Host-Range Inducible Expression Vector

通用定義 / General Definition:

BioVector? pSRK-GM 是一種基于 BHR（Broad-Host-Range） 復(fù)制子的廣宿主表達(dá)載體，屬于著名的 pSRK 系列。該載體利用 $lacI^q$ - $P_{lac}$ 誘導(dǎo)系統(tǒng)，能夠在多種革蘭氏陰性菌（如假單胞菌 Pseudomonas、根瘤菌 Rhizobium、農(nóng)桿菌 Agrobacterium 等）中實(shí)現(xiàn)基因的嚴(yán)謹(jǐn)調(diào)控表達(dá)。其名稱中的 "GM" 代表其攜帶 慶大霉素（Gentamicin） 抗性標(biāo)記，這使其在那些對氨芐青霉素或卡那霉素具有天然抗性的環(huán)境分離株中極具應(yīng)用價(jià)值。

BioVector? pSRK-GM is a broad-host-range (BHR) expression vector based on the pSRK series. It utilizes the $lacI^q$ - $P_{lac}$ system to achieve tightly regulated gene expression in a wide variety of Gram-negative bacteria, including Pseudomonas, Rhizobium, and Agrobacterium. The "GM" designation refers to its Gentamicin resistance marker, making it an invaluable tool for genetic studies in environmental isolates that possess intrinsic resistance to ampicillin or kanamycin.

BioVector? pSRK-GM 載體技術(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? pSRK-GM 表達(dá)載體

• 載體類型： 廣宿主誘導(dǎo)型表達(dá)載體

• 復(fù)制子：$BHR$ (通?；?$pBBR1$ 來源，具有高度相容性)

• 誘導(dǎo)系統(tǒng)： IPTG 誘導(dǎo) ($lacI^q$ 控制下的 $P_{lac}$ 啟動子)

• 抗性基因： 慶大霉素 (Gentamicin)

• 克隆方法： 限制性內(nèi)切酶克隆 (多克隆位點(diǎn)含 $EcoRI, KpnI, BamHI$ 等)

2. 培養(yǎng)與擴(kuò)增條件

• 大腸桿菌擴(kuò)增： 使用 BioVector? LB 培養(yǎng)基，添加 10-15 $\mu$g/mL BioVector? 慶大霉素，37°C 培養(yǎng)。

• 廣宿主轉(zhuǎn)化： 可通過接合轉(zhuǎn)移 (Conjugation) 或電穿孔法導(dǎo)入目標(biāo)菌株。

• 培養(yǎng)環(huán)境： 根據(jù)目標(biāo)菌株的最適溫度設(shè)定（通常為 28°C - 30°C 用于環(huán)境菌株）。

3. 載體應(yīng)用

• 跨物種基因表達(dá)： 在非模式革蘭氏陰性菌中驗(yàn)證基因功能。

• 互補(bǔ)實(shí)驗(yàn)： 用于細(xì)菌突變株的功能恢復(fù)實(shí)驗(yàn)。

• 環(huán)境微生物學(xué)： 研究土壤或水生細(xì)菌在誘導(dǎo)條件下的代謝路徑。

4. 注意事項(xiàng)

• 背景表達(dá)：$P_{lac}$ 在某些物種中可能存在一定的背景漏表達(dá) (Leaky expression)，若表達(dá)毒性蛋白需謹(jǐn)慎。

• 質(zhì)粒穩(wěn)定性： 在沒有抗性壓力的情況下，部分 BHR 質(zhì)?？赡軙G失，建議持續(xù)維持 BioVector? 慶大霉素選擇壓力。

English Datasheet

1. General Product Information

• Product Name: BioVector? pSRK-GM Expression Vector

• Vector Type: Broad-Host-Range Inducible Expression Vector

• Replicon:$BHR$ (derived from $pBBR1$, ensuring broad compatibility)

• Induction System: IPTG-inducible ($P_{lac}$ promoter controlled by $lacI^q$)

• Antibiotic Resistance: Gentamicin

• Cloning Method: Restriction Enzyme Cloning (MCS includes $EcoRI, KpnI, BamHI$, etc.)

2. Culture and Amplification Conditions

• E. coli Propagation: Use BioVector? LB Medium supplemented with 10-15 $\mu$g/mL BioVector? Gentamicin, incubated at 37°C.

• Host Transformation: Delivery via conjugation (triparental mating) or electroporation into target species.

• Incubation: Set according to the optimal growth temperature of the host (typically 28°C - 30°C for environmental microbes).

3. Applications

• Cross-species Gene Expression: Validating gene function in non-model Gram-negative bacteria.

• Complementation Assays: Restoring phenotype in bacterial mutant strains.

• Environmental Microbiology: Investigating metabolic pathways in soil or aquatic bacteria under induced conditions.

4. Key Usage Notes

• Basal Expression: The $P_{lac}$ promoter may exhibit "leaky" expression in certain species; exercise caution when expressing proteins toxic to the host.

• Plasmid Stability: Some BHR plasmids can be lost without selective pressure. It is recommended to maintain BioVector? Gentamicin selection throughout the experiment.

Note: For efficient transfer into environmental strains, BioVector? suggests using the $E. coli$ S17-1 or WM3064 donor strains for conjugation. Always verify the plasmid structure after mobilization using BioVector? Plasmid Mini Kits.

Supplier生產(chǎn)廠家: BioVector NTCC典型培養(yǎng)物保藏中心

E-mail: [email protected]

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