Himar1 mariner/Tc1 BioVector® plasmid转座子质粒载体

BioVector® Himar1 is a synthetic DNA transposon derived from the horn fly (Haematobia irritans). It is a member of the mariner/Tc1 family and functions through a cut-and-paste mechanism that targets TA dinucleotides for insertion.

Functional Properties

• Target Specificity: It integrates exclusively at TA dinucleotides. Upon insertion, the TA sequence is duplicated at both ends of the transposon.

• Host Independence: Unlike many other systems, Himar1 does not require specific host-encoded proteins for transposition, making it a universal tool for mutagenesis in bacteria, fungi, and animal cells.

• Hyperactive Variants: Standard Himar1 has low activity. Researchers typically use hyperactive mutants like C9 or A7, which offer significantly higher integration rates.

• ITRs (Inverted Terminal Repeats): These are the 27-28 bp sequences flanking the DNA to be moved. The transposase recognizes these sequences to initiate the "cut."

• Transposase Gene: Often placed under the control of an inducible promoter (such as Pxylose or Ptet) to prevent continuous jumping and genomic instability.

• Selectable Markers: The transposon "cargo" usually includes an antibiotic resistance gene (e.g., Kanamycin or Erythromycin) to identify cells where successful genomic integration has occurred.

• Counter-Selection: Many Himar1 plasmids use temperature-sensitive origins of replication. This allows the researcher to "cure" the plasmid from the cell population by raising the growth temperature after transposition is complete.

Available Plasmid Resources

• pSAM_Ec (Hyperactive C9): A widely used version for high-efficiency mutagenesis in various bacterial species, available at BioVector #102939.

• pDonor_Tn (Himar1C9): A modern vector using anhydrotetracycline (ATc) induction for controlled transposition, available at BioVector #213906.

• pCis mCherry-SS (Hyperactive A7): Specialized for use in difficult-to-transform species like Rickettsia, providing a fluorescent marker for tracking, available at BioVector #84687.

• 名称： Himar1 (源自角蝇 Haematobia irritans)

• 家族：mariner/Tc1 超家族，II 类 DNA 转座子

• 机制： “剪切-粘贴”（Cut-and-paste）式保守转座

• 目标位点： 严格特异于 TA 二核苷酸，并在插入处产生 TA 序列重复

• 宿主独立性： Himar1 转座不依赖于特定的宿主因子，因此广泛适用于细菌（革兰氏阴性/阳性）、真菌、昆虫甚至哺乳动物细胞

• 高活性突变体： 常用的超活跃型变体包括 C9 和 A7。C9 突变体（如在 pDonor_Tn 质粒中）可将转座效率提高 4 至 50 倍

• 诱导系统： 现代载体（如 pBLAM1-x 系列）通常配备 ATc (脱水四环素) 或木糖诱导启动子，以精确控制转座酶的表达时间

• ITR (反向终端重复序列)： 长度约 27-28 bp，是转座酶识别并切割的边界

• 抗性标记： 常见的载体携带 Kanamycin (卡那霉素)、Chloramphenicol (氯霉素) 或 Erythromycin (红霉素) 用于筛选基因组整合成功的克隆

• 温度敏感型复制子： 许多质粒（如 repA(ts)）在高温（如 40°C）下无法复制，便于在转座发生后通过“质粒消除”（Plasmid curing）获得纯净的基因组突变株

1. 随机诱变库构建： 产生高复杂度的插入突变库，用于功能基因组学研究

2. 基因交付： 将外源基因（如荧光蛋白 mCherry）稳定整合进宿主染色体

3. Tn-Seq 库制备： 结合 MmeI 等限制性内切酶位点，支持高通量转座子测序

• BioVector 资源：

  • pDonor_Tn[himar1C9] (ID: 213906) - ATc 诱导型。

  • pSAM_Ec (ID: 102939) - 经典的细菌随机突变载体。

  • pCis mCherry-SS (ID: 84687) - 带 mCherry 标记的 A7 变体。

  
  

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