BioVector? NTCC? pSC189細(xì)菌轉(zhuǎn)座子遞送載體BioVector? C9轉(zhuǎn)座酶質(zhì)粒

pSC189 BioVector?質(zhì)粒是一種細(xì)菌轉(zhuǎn)座子遞送載體，用于隨機(jī)插入誘變，特別是結(jié)合 mariner Himar1 轉(zhuǎn)座子系統(tǒng)。
主要特性和信息?特性?詳情載體類(lèi)型轉(zhuǎn)座子遞送（自殺）載體大小6982 堿基對(duì) (bp)宿主生物大腸桿菌（需要 $\lambda$pir 溶原宿主，如 SM10 $\lambda$pir 或 PIR1，才能復(fù)制）細(xì)菌抗性氨芐青霉素 (100 μg/mL) 和卡那霉素 (50 μg/mL)復(fù)制起點(diǎn)R6K \(\gamma \) 復(fù)制起點(diǎn) ($ori$R6K) 和 RP4 轉(zhuǎn)移起點(diǎn) (\(oriT\))拷貝數(shù)低拷貝關(guān)鍵組件包含高活性的 Himar1 C9 轉(zhuǎn)座酶基因、T3 和 T7 啟動(dòng)子以及 3xFLAG 表位標(biāo)簽研究中的應(yīng)用?pSC189 質(zhì)粒在接合實(shí)驗(yàn)中被用作“自殺載體”。因?yàn)樗?R6K 復(fù)制起點(diǎn)，所以它只能在表達(dá) $\lambda$pir 蛋白的大腸桿菌菌株中復(fù)制。當(dāng)轉(zhuǎn)移到缺乏這種蛋白的受體細(xì)菌物種中時(shí)，該質(zhì)粒無(wú)法復(fù)制。然后，轉(zhuǎn)座子（由 pSC189 主鏈攜帶）從非復(fù)制質(zhì)粒跳入宿主基因組，從而實(shí)現(xiàn)穩(wěn)定且可遺傳的隨機(jī)基因插入。?隨機(jī)誘變：用于通過(guò)將轉(zhuǎn)座子隨機(jī)插入各種革蘭氏陰性細(xì)菌（包括弧菌屬和鉤端螺旋體屬物種）的基因組中來(lái)生成大型突變體庫(kù)?；虬l(fā)現(xiàn)：篩選所得的文庫(kù)，以鑒定必需基因或參與特定過(guò)程（如毒力、應(yīng)激抗性或核酸酶活性）的基因。表位標(biāo)記：該載體旨在促進(jìn)使用 3xFLAG 表位標(biāo)簽標(biāo)記蛋白質(zhì)，有助于功能研究和蛋白質(zhì)表征。
The BioVector? pSC189 plasmid is a bacterial transposon delivery vector used for random insertional mutagenesis, particularly with the mariner Himar1 transposon system.
Key Features and Information?Feature?DetailVector TypeTransposon delivery (suicide) vectorSize6982 base pairs (bp)Host OrganismE. coli (requires a $\lambda$pir lysogen host like SM10 $\lambda$pir or PIR1 for replication)Bacterial ResistanceAmpicillin (100 μg/mL) and Kanamycin (50 μg/mL)Origin of ReplicationR6K \(\gamma \) origin of replication ($ori$R6K) and RP4 origin of transfer (\(oriT\))Copy NumberLow CopyKey ComponentsContains the hyperactive Himar1 C9 transposase gene, T3 and T7 promoters, and 3xFLAG epitope tagUsage in Research?The pSC189 plasmid is used as a "suicide vector" in conjugation experiments. Because it contains the R6K origin of replication, it can only replicate in E. coli strains that express the $\lambda$pir protein. When transferred into a recipient bacterial species that lacks this protein, the plasmid cannot replicate. The mariner transposon (which is carried on the pSC189 backbone) then jumps from the non-replicating plasmid into the host's genome, allowing for random gene insertions that are stable and heritable.?Random Mutagenesis: Used to generate large libraries of mutants by random insertion of the transposon into the genome of various Gram-negative bacteria, including Vibrio and Leptospira species.Gene Discovery: The resulting libraries are screened to identify essential genes or genes involved in specific processes like virulence, stress resistance, or nuclease activity.Epitope Tagging: The vector is designed to facilitate the tagging of proteins with a 3xFLAG epitope tag, aiding in functional studies and protein characterization.


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