快速赛车骗局:內部核糖體進入位點(IRES) — 實現同一載體高效共表達兩個基因的元件

簡介

 IRES是實現同一載體高效共表達兩個基因的元件。一般可以通過裝載熒光標簽或可以在表達的同時生物素化的蛋白到同一載體來檢測一個基因的轉染效率。IRES元件可以用于以下研究:

  • 檢測基因轉染效率 
  • 檢測蛋白修飾 
  • 體內生物素化 
  • 穩定轉染 

 

OmicsLink ORF cDNA clones with IRES element

 

技術

IRES技術可以使用同一啟動子在同一載體高效共表達兩個基因的元件 (見figures 1和 2)。GeneCopoeia已經構建了許多包含IRES元件的載體。

 

Combined with OmicsLink expression-ready clones

為了滿足不同需求,GeneCopoeia已經將IRES元件構建到 部分哺乳動物細胞和慢病毒載體,有以下報告基因可?。?/p>

  • eGFP
  • mCherry
  • SUMO
  • 生物素連接酶
  • 新霉素
  • 熒光素酶

 

 GeneCopeia提供帶有IRES元件、CMV 啟動子的20,000多種人和15,000多種小鼠的表達克隆。

Choice of co-expressing any reporter gene with IRES element

 

優勢

  • 可以將報告基因對靶蛋白的生物學活性的影響降到最低
  • 可以有效共轉染因為兩個基因在同一載體

 

 

 

 

 

OmicsLink ORF cDNA clones with IRES element

 

 

 

 

圖1. 帶有IRES元件的OmicsLink™ 即用型ORF表達表達克隆

應用實例

應用

  • 使報告基因如 eGFP、mCherry 或 Luciferase與靶蛋白共表達以檢測轉染效率
  • 可以通過共表達如SUMO這樣特定的蛋白修飾酶來監測蛋白修飾
  • 共表達生物素連接酶可以在體內生物素化帶有AviTag的蛋白
  • 共表達如Neomycin這樣的篩選標記可以進行穩定細胞株的篩選

 

嚴格的質檢和質量保證

OmicsLink clone collection里的所有ORF表達克隆源于驗證過的全長cDNA克隆或高質量的人源cDNA組織文庫。

寄送之前,GeneCopoeia對所有的克隆進行了嚴格的質檢過程。

  • 所有的ORF經過全長測序
  • 通過PCR驗證序列長度
  • 通過酶切驗證質粒的正確性

IRES作用流程

Schematic of protein expression with IRES technology

Specific biotinylation of AviTag eGFP by E. coli biotin ligase in 293 cells

圖2. 在293細胞通過大腸桿菌生物素連接酶特異性的生物素化AviTag

載體信息

使用IRES技術的表達載體列表

載體 啟動子 宿主細胞 篩選標記 標簽 蛋白酶位點
pReceiver-Lv165 EF1a Lentiviral N/A IRES2-eGFP N/A
pReceiver-Lv166 EF1a Lentiviral N/A IRES2-mcherry N/A
pReceiver-Lv201 CMV Lentiviral Puromycin SV40-eGFP-IRES-puromycin N/A
pReceiver-Lv202 CMV Lentiviral Puromycin C-Myc-SV40-eGFP-IRES-puromycin N/A
pReceiver-Lv203 CMV Lentiviral Puromycin C-Flag-SV40-eGFP-IRES-puromycin N/A
pReceiver-Lv204 CMV Lentiviral Puromycin C-Myc-SV40-mCherry-IRES-puromycin N/A
pReceiver-Lv205 CMV Lentiviral Puromycin IRES2-eGFP-IRES-puromycin N/A
pReceiver-Lv206 CMV Lentiviral Puromycin SV40-mCherry-IRES-puromycin N/A
pReceiver-Lv207 CMV Lentiviral Hygromycin SV40-eGFP-IRES-hygromycin N/A
pReceiver-Lv211 CMV Lentiviral Hygromycin C-HaloTag-IRES-hygromycin N/A
pReceiver-Lv213 CMV Lentiviral Puromycin IRES2-mCherry-IRES-puromycin N/A
pReceiver-Lv214 CMV Lentiviral N/A IRES2-mCherry N/A
pReceiver-Lv215 CMV Lentiviral N/A IRES2-eGFP N/A
pReceiver-Lv216 CMV Lentiviral Puromycin C-Flag-SV40-mCherry-IRES-puromycin N/A
pReceiver-Lv217 CMV Lentiviral Puromycin SV40-hLUC-IRES-puromycin N/A
pReceiver-Lv218 CMV Lentiviral Neomycin IRES2-Neomycin N/A
pReceiver-Lv219 CMV Lentiviral Puromycin IRES2-Puromycin N/A
pReceiver-Lv220 CMV Lentiviral Hygromycin IRES2-Hygromycin N/A
pReceiver-Lv221 CMV Lentiviral N/A IRES2-eYFP N/A
pReceiver-Lv222 CMV Lentiviral N/A IRES2-eCFP N/A
pReceiver-Lv224 EF1a Lentiviral Puromycin IRES2-mcherry-IRES-puromycin N/A
pReceiver-Lv225 EF1a Lentiviral Puromycin IRES2-eGFP-IRES-puromycin N/A
pReceiver-Lv228 PGK Lentiviral N/A IRES2-eGFP N/A
pReceiver-Lv229 PGK Lentiviral N/A IRES2-mcherry N/A
pReceiver-Lv230 PGK Lentiviral Puromycin IRES2-mcherry-IRES-puromycin N/A
pReceiver-Lv231 PGK Lentiviral Puromycin IRES2-eGFP-IRES-puromycin N/A
pReceiver-Lv233 EF1a Lentiviral Puromycin SV40-eGFP-IRES-Puromycin N/A
pReceiver-Lv234 CAG Lentiviral N/A IRES2-eGFP N/A
pReceiver-Lv235 CAG Lentiviral Puromycin SV40-eGFP-IRES-Puromycin N/A
pReceiver-Lv244 CMV Lentiviral Hygromycin SV40-mCherry-IRES-Hygromycin N/A
pReceiver-Lv245 EF1a Lentiviral Puromycin cMyc-SV40-mCherry-IRES-Puromycin N/A
pReceiver-Lv246 UBC Lentiviral Puromycin SV40-eGFP-IRES-Puromycin N/A
pReceiver-Lv247 UBC Lentiviral Puromycin IRES2-mCherry-IRES-Puromycin N/A
pReceiver-M45 CMV Mammalian Neomycin C-3xHA+IRES-eGFP N/A
pReceiver-M46 CMV Mammalian Neomycin C-Flag+IRES-eGFP N/A
pReceiver-M48 CMV Mammalian Neomycin N-Avi+IRES-Biotin ligase N/A
pReceiver-M51 CMV Mammalian Neomycin C-His+IRES-eGFP N/A
pReceiver-M61 CMV Mammalian Neomycin IRES2-eGFP N/A
pReceiver-M62 CMV Mammalian Neomycin C-Avi-IRES-Biotin ligase N/A
pReceiver-M72 CMV Mammalian Neomycin C-Myc+IRES2-eGFP N/A
pReceiver-M73 CMV Mammalian Neomycin C-Myc-IRES2-mCherry N/A
pReceiver-M83 CMV Mammalian Neomycin IRES2-mCherry N/A
pReceiver-M90 CMV Mammalian Puromycin SV40-eGFP-IRES-Puromycin N/A
pReceiver-M91 CMV Mammalian Puromycin C-3xHA-SV40-eGFP-IRES-Puromycin N/A
pReceiver-M92 CMV Mammalian Puromycin C-Myc-SV40-eGFP-IRES-Puromycin N/A
pReceiver-M93 CMV Mammalian Puromycin C-Flag-SV40-eGFP-IRES-Puromycin N/A
pReceiver-M95 CMV Mammalian Puromycin SV40-mCherry-IRES-Puromycin N/A

 

點擊了解即用型ORF克隆 ,提供大腸桿菌、哺乳動物細胞、昆蟲細胞、酵母和麥胚無細胞表達克隆載體和50種融合標簽。

 

 

參考文獻

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