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.

• 名稱： Himar1 (源自角蠅 Haematobia irritans)

• 家族：mariner/Tc1 超家族，II 類 DNA 轉(zhuǎn)座子

• 機制： “剪切-粘貼”（Cut-and-paste）式保守轉(zhuǎn)座

• 目標位點： 嚴格特異于 TA 二核苷酸，并在插入處產(chǎn)生 TA 序列重復(fù)

• 宿主獨立性： Himar1 轉(zhuǎn)座不依賴于特定的宿主因子，因此廣泛適用于細菌（革蘭氏陰性/陽性）、真菌、昆蟲甚至哺乳動物細胞

• 高活性突變體： 常用的超活躍型變體包括 C9 和 A7。C9 突變體（如在 pDonor_Tn 質(zhì)粒中）可將轉(zhuǎn)座效率提高 4 至 50 倍

• 誘導(dǎo)系統(tǒng)： 現(xiàn)代載體（如 pBLAM1-x 系列）通常配備 ATc (脫水四環(huán)素) 或木糖誘導(dǎo)啟動子，以精確控制轉(zhuǎn)座酶的表達時間

• ITR (反向終端重復(fù)序列)： 長度約 27-28 bp，是轉(zhuǎn)座酶識別并切割的邊界

• 抗性標記： 常見的載體攜帶 Kanamycin (卡那霉素)、Chloramphenicol (氯霉素) 或 Erythromycin (紅霉素) 用于篩選基因組整合成功的克隆

• 溫度敏感型復(fù)制子： 許多質(zhì)粒（如 repA(ts)）在高溫（如 40°C）下無法復(fù)制，便于在轉(zhuǎn)座發(fā)生后通過“質(zhì)粒消除”（Plasmid curing）獲得純凈的基因組突變株

1. 隨機誘變庫構(gòu)建： 產(chǎn)生高復(fù)雜度的插入突變庫，用于功能基因組學(xué)研究

2. 基因交付： 將外源基因（如熒光蛋白 mCherry）穩(wěn)定整合進宿主染色體

3. Tn-Seq 庫制備： 結(jié)合 MmeI 等限制性內(nèi)切酶位點，支持高通量轉(zhuǎn)座子測序

• BioVector 資源：

  • pDonor_Tn[himar1C9] (ID: 213906) - ATc 誘導(dǎo)型。

  • pSAM_Ec (ID: 102939) - 經(jīng)典的細菌隨機突變載體。

  • pCis mCherry-SS (ID: 84687) - 帶 mCherry 標記的 A7 變體。

  
  

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