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Home > ÀüÁ¦Ç°º¸±â > Inducible System ¡¤ Yeast hybrid > Tet À¯µµ ¹ßÇö ½Ã½ºÅÛ > [Legacy] Tetracycline À¯µµ¹ßÇö ½Ã½ºÅÛ

[Legacy] Tetracycline À¯µµ¹ßÇö ½Ã½ºÅÛ

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Clontech
631070
pTet-Off Advanced Vector
°ü·ÃÇмú ±¸¸ÅÇϱ⠶óÀ̼±½º 
20 ¥ìg
818,400¿ø 
1,023,000¿ø
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11.01 ~ 12.27
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x33574
Clontech
631059
pTRE-Tight Vector
°ü·ÃÇмú ±¸¸ÅÇϱ⠶óÀ̼±½º 
20 ¥ìg
681,600¿ø 
852,000¿ø
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11.01 ~ 12.27
Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â
x33606, x33152
Clontech
632105
Retro-X Tet-Off Advanced Inducible Expression System
°ü·ÃÇмú ±¸¸ÅÇϱ⠶óÀ̼±½º 
1 Kit
(Package)
1,545,600¿ø 
1,932,000¿ø
°¡°ÝÇÒÀÎ
11.01 ~ 12.27
Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â
x33549, x33555, x33556, x32537, x33013
Clontech
632163
Lenti-X¢â Tet-Off¢ç Advanced Inducible Expression System
°ü·ÃÇмú ±¸¸ÅÇϱ⠶óÀ̼±½º 
1 Set
(Package)
2,632,000¿ø 
3,290,000¿ø
°¡°ÝÇÒÀÎ
11.01 ~ 12.27
Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â
x33474, x32766, x32768, x32774, x32784, x33200
Clontech
631311
Doxycycline
°ü·ÃÇмú ±¸¸ÅÇϱâ
5 g
300,800¿ø 
376,000¿ø
°¡°ÝÇÒÀÎ
11.01 ~ 12.27
Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â
x94899, x32770, x32771

  • ³ôÀº À¯µµ¹ßÇöÀ²°ú ±Øµµ·Î ³·Àº basal expression
  • cDNA¿Í miRNAÀÇ °í°¨µµ ¹ßÇö Á¶Àý
  • Plasmid, lentivirus, retroivirus ÇüÅ·Π´Ù¾çÇÏ°Ô ÀÌ¿ë °¡´É
Tet-On Advanced¿Í Tet-Off Advanced Inducible Gene Expression SystemÀº E. coli ÀÇ Á¶Àý ±âÀÛÀÎ tetracycline-resistance operon¿¡ ±â¹ÝÀ» µÎ°í ÀÖÀ¸¸ç, °¢ systemÀº Æ÷À¯·ù ¼¼Æ÷¿¡ Àû¿ëÇÒ ¼ö ÀÖµµ·Ï ÃÖÀûÈ­µÈ 2 °³ÀÇ ±¸¼º¿ä¼Ò·Î µÇ¾î ÀÖ´Ù (Figure 1):
  • Regulator vector´Â tetracyclineÀ¸·Î Á¶ÀýµÇ´Â Tet-On Advanced ¶Ç´Â Tet-Off Advanced transactivator¸¦ ¹ßÇöÇÏ´Â vectorÀÌ´Ù. ÀÌ transactivator´Â Á¶Àý ÀÎÀÚÀÎ doxycycline (Dox ; tetracycline À¯µµÃ¼)¿¡ ´ëÇÏ¿© ¹Ý´ë·Î ¹ÝÀÀÇϸç, 3 °³ÀÇ minimal VP16 transcription activation domainÀ» ¿¬°áÇÏ¿© ÀçÁ¶ÇÕÇÑ bacterial Tet Repressor (TetR)·Î ±¸¼ºÀÌ µÇ¾î ÀÖ´Ù (1).
  • Response vectorÀÎ pTRE-Tight´Â ¸ñÀû À¯ÀüÀÚÀÇ ¹ßÇöÀ» Á¶ÀýÇÏ´Â promoter¿¡ ±â´ÉÀÌ Çâ»óµÈ tetracycline response element (TRE)¸¦ °¡Áö°í ÀÖ´Ù.
ÀÌ ½Ã½ºÅÛÀ¸·Î ±¸ÃàµÈ cell line¿¡¼­´Â ¸ñÀû À¯ÀüÀÚÀÇ ¹ßÇö Á¤µµ¸¦ DoxÀÇ Ã·°¡·®¿¡ µû¶ó Á¤È®ÇÏ°Ô Á¶ÀýÇÒ ¼ö ÀÖ´Ù (Figure 2). Tet-Advanced SystemÀÇ ÃÖ´ë ¹ßÇö ¼öÁØÀº ¾ÆÁÖ ³ô°í ƯÀÌÀûÀÏ »Ó ¾Æ´Ï¶ó Dox ÀÌ¿ÜÀÇ ¼¼Æ÷ ³» ÀÎÀÚ¿¡ ÀÇÇÏ¿© ¿µÇâÀ» ¹ÞÁö ¾Ê´Â´Ù. Clontech¿¡¼­´Â ¹Ì¸® Á¦ÀÛµÈ Tet-Advanced Cell LineÀ» ÆǸÅÇÏ°í ÀÖ´Ù.
  • Tet-On Advanced transactivator´Â DoxÀÇ Á¸Àç ÇÏ¿¡¼­ TRE promoter¿¡ °áÇÕÇÏ¿© À¯ÀüÀÚ ¹ßÇöÀ» È°¼ºÈ­½ÃŲ´Ù.
  • Tet-Off Advanced transactivator´Â Dox°¡ ¾øÀ» °æ¿ì TRE promoter¿¡ °áÇÕÇÏ¿© À¯ÀüÀÚ ¹ßÇöÀ» È°¼ºÈ­½ÃŲ´Ù.
ClontechÀº 2 Á¾·ùÀÇ ¾ÆÁÖ È¿À²ÀûÀÎ virus µµÀÔ¿ë Tet-advanced systemÀÎ Retro-X Tet-Advanced Inducible Expression System°ú Lenti-X Tet-Advanced Inducible Expression SystemÀÇ À» ÆǸÅÇÏ°í ÀÖÀ¸¸ç ´ëºÎºÐÀÇ ¼¼Æ÷¿¡ Àû¿ëÀÌ °¡´ÉÇÏ´Ù. ƯÈ÷, Lenti-X lentivirus systemÀº primary culture, Áٱ⼼Æ÷, ½Å°æ¼¼Æ÷, ºñºÐ¿­¼¼Æ÷¿¡ À¯ÀüÀÚ¸¦ µµÀÔÇÒ °æ¿ì ¸Å¿ì À¯¿ëÇϸç, ³ôÀº ¿ª°¡ (titer)¸¦ º¸ÀåÇÏ´Â Lenti-X HT Packaging SystemÀ» Æ÷ÇÔÇÏ°í ÀÖ´Ù.


Figure 1. Induced expression in the Tet-Off Advanced and Tet-On Advanced Systems. The Tet-controlled transactivators are fusion proteins that contain a TetR-derived DNA-binding domain joined to three minimal transcription activation domains from HSV VP16. Each transactivator has been optimized for expression in mammalian cells.


Figure 2. Induced luciferase expression in Tet-On Advanced and Tet-Off Advanced Expression Systems. In the absence of Dox, basal expression is undetectable in Tet-On Advanced cells, whereas full induction in the presence of Dox is very high. Tet-Off Advanced cells show extremely high induction in the absence of Dox, and undetectable basal expression in its presence.
Components & Storage Conditions
°¢ Á¦Ç° ±¸¼º¹°°ú º¸°üÁ¶°ÇÀº Certificate of Analysis ¸¦ ÂüÁ¶ÇϽʽÿÀ.
References
1. Urlinger, S. et al. (2000) Proc. Natl. Acad. Sci. USA 97(14):7963-7968.

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