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 Á¦Ç°¾È³»  Ç×ü,Ç׿ø,ŶƮ   Cancer Research  Topoisomerase Assay

Bacterial DNA Gyrase Assay Kit

´ëÀå±Õ, Ȳ»öÆ÷µµ»ó±¸±ÕÀ¸·ÎºÎÅÍ Á¤Á¦ÇÑ Bacterial DNA Gyrase¿Í À̵éÀ» ½Å¼ÓÇÏ°Ô ºÐ¼®Çϱâ À§ÇÑ Kit¸¦ ¼Ò°³ÇÕ´Ï´Ù.

TopoGEN
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  • E. coli Bacterial DNA Gyrase
    • in vitro¿¡¼­ÀÇ Plasmid supercoiling ºÐ¼® ȤÀº ½Å¾à ½ºÅ©¸®´×À» À§ÇÑ Á¦Ç°ÀÔ´Ï´Ù.
    • °íµµ·Î Á¤Á¦µÈ DNA gyrase´Â A, B subunitsÀÌ ¸ðµÎ Æ÷ÇÔµÈ Holoenzyme ÇüÅ·ΠÁ¦°øµË´Ï´Ù.
    • 5X Assay buffer¿Í Dilution buffer°¡ ÇÔ²² Á¦°øµÇ¸ç, º°µµ ±¸¸Å °¡´ÉÇÕ´Ï´Ù.
    • 1 UnitÀÇ Gyrase´Â 37°C¿¡¼­ 30ºÐ ³» 0.2 ugÀÇ Plasmid¸¦ supercoiling ÇÕ´Ï´Ù.
    Figure 1. Purification of Gyrase A and B Subunits
  • S. Aureus DNA Gyrase
    • ±×¶÷¾ç¼º¼¼±ÕÀΠȲ»öÆ÷µµ»ó±¸±ÕÀ¸·ÎºÎÅÍ Á¤Á¦µÈ DNA gyrase·Î½á ½Å¾à °³¹ß, ½ºÅ©¸®´× µî ¸ðµç Ãø¸é¿¡¼­ Àû¿ëÀÌ °¡´ÉÇÕ´Ï´Ù.
    • kDNAÀÇ Decatenating°ú SupercoilingÀ» Ȱ¼ºÈ­ ½Ãų »Ó¸¸ ¾Æ´Ï¶ó Relaxed plasmid DNA¸¦ Negatively supercoiling ÇÕ´Ï´Ù.
    Figure 2. Assays should be performed in a final volume of 20 ul by adding appropriate amounts of buffers, DNA enzyme
  • E. coli DNA Gyrase and Relaxed DNA Assay Kit
    • DNA gyraseÀ» ½Å¼ÓÇϰí Á¤È®ÇÏ°Ô ºÐ¼®ÇÒ ¼ö ÀÖµµ·Ï µðÀÚÀεǾú½À´Ï´Ù.
    • Controlled reagent°¡ Á¦°øµÇ¾î GyraseÀÇ Á¤Á¦¿Í ºÐ¼®¿¡ ´ëÇÑ Áõ¸íÀÌ °¡´ÉÇϸç, »õ·Î¿î anti-gyrase Ȱ¼º ¾à¹°À» ´ë·®À¸·Î ½ºÅ©¸®´×ÇÒ ¶§ ÀûÇÕÇÑ Á¦Ç°ÀÔ´Ï´Ù.
    • Kit ±¸¼ºÇ°
      • Relaxed pHOT-1 DNA
      • High purity E. coli DNA gyrase enzyme
      • Assay buffer
      • Dilution buffer for enzyme
  • Gyrase Assay Kit
      • »õ·Ó°Ô °³¹ßÇÑ Gel systemÀ» »ç¿ëÇÏ¿© ¸Å¿ì ½Å¼ÓÇÏ°í ¸íÈ®ÇÏ°Ô Topoisomerase II È¿¼Ò (DNA Gyrase) Ȱ¼º ÃøÁ¤ÀÌ °¡´ÉÇÕ´Ï´Ù.
      • Topo II È¿¼ÒÀÇ ±âº» ºÐ¼®À» À§ÇØ ÇÊ¿äÇÑ Buffer, DNA substrare, DNA marker°¡ Á¦°øµÇ¸ç, EnzymeÀº Á¦°øµÇÁö ¾Ê½À´Ï´Ù.
      • °£´ÜÇÑ Ã˸ÅȰ¼º ¾ïÁ¦ ¾à¹° »Ó¸¸ ¾Æ´Ï¶ó Gyrase¿¡ ÀÇÇÑ DNA Àý´ÜÀ» ÀÚ±ØÇÏ´Â µ¶±Ø¹°À» °¨ÁöÇÕ´Ï´Ù.
      • kDNA ±â¹ÝÀÇ Gyrase ºÐ¼®¹æ¹ýÀº Crude mixture ȤÀº ÃßÃâ¹°·ÎºÎÅÍ ½±°í ºü¸£°Ô Inhibitor¸¦ Á¤Á¦ÇÒ ¼ö ÀÖ´Â ÀåÁ¡ÀÌ ÀÖ½À´Ï´Ù.
      • Kit ±¸¼ºÇ°
        • Kinetoplast DNA (kDNA)
        • Marker linear kDNA
        • Marker decatenated kDNA (topo II treated)
        • 10X Gyrase assay buffer
        • Gyrase stop and load buffer
      • ºÐ¼® ¹æ¹ý




    Figure 3. Interpretation of Gels