BioVector® pSRK-GM 广宿主诱导型表达载体 / BioVector® pSRK-GM Broad-Host-Range Inducible Expression Vector

通用定义 / General Definition:

BioVector® pSRK-GM 是一种基于 BHR（Broad-Host-Range） 复制子的广宿主表达载体，属于著名的 pSRK 系列。该载体利用 $lacI^q$ - $P_{lac}$ 诱导系统，能够在多种革兰氏阴性菌（如假单胞菌 Pseudomonas、根瘤菌 Rhizobium、农杆菌 Agrobacterium 等）中实现基因的严谨调控表达。其名称中的 "GM" 代表其携带 庆大霉素（Gentamicin） 抗性标记，这使其在那些对氨苄青霉素或卡那霉素具有天然抗性的环境分离株中极具应用价值。

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 载体技术说明书 (Plasmid Datasheet)

中文版说明书

1. 产品基本信息

• 产品名称： BioVector® pSRK-GM 表达载体

• 载体类型： 广宿主诱导型表达载体

• 复制子：$BHR$ (通常基于 $pBBR1$ 来源，具有高度相容性)

• 诱导系统： IPTG 诱导 ($lacI^q$ 控制下的 $P_{lac}$ 启动子)

• 抗性基因： 庆大霉素 (Gentamicin)

• 克隆方法： 限制性内切酶克隆 (多克隆位点含 $EcoRI, KpnI, BamHI$ 等)

2. 培养与扩增条件

• 大肠杆菌扩增： 使用 BioVector® LB 培养基，添加 10-15 $\mu$g/mL BioVector® 庆大霉素，37°C 培养。

• 广宿主转化： 可通过接合转移 (Conjugation) 或电穿孔法导入目标菌株。

• 培养环境： 根据目标菌株的最适温度设定（通常为 28°C - 30°C 用于环境菌株）。

3. 载体应用

• 跨物种基因表达： 在非模式革兰氏阴性菌中验证基因功能。

• 互补实验： 用于细菌突变株的功能恢复实验。

• 环境微生物学： 研究土壤或水生细菌在诱导条件下的代谢路径。

4. 注意事项

• 背景表达：$P_{lac}$ 在某些物种中可能存在一定的背景漏表达 (Leaky expression)，若表达毒性蛋白需谨慎。

• 质粒稳定性： 在没有抗性压力的情况下，部分 BHR 质粒可能会丢失，建议持续维持 BioVector® 庆大霉素选择压力。

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生产厂家: BioVector NTCC典型培养物保藏中心

E-mail: BioVector@163.com

http://www.biovector.net