¡Ø º» Á¦Ç°Àº CloningÀ» À§ÇØ In-Fusion
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Á¦Ç° »ç¿ë Àü In-Fusion¢ç ¿ø¸®(µ¿¿µ»ó¸µÅ©)¸¦ ¹Ýµå½Ã Âü°íÇØÁֽñ⠹ٶø´Ï´Ù.
¢º º» Á¦Ç°Àº 2¼¼´ë ¹öÀüÀÇ Tetracycline-inducible expression system ÀÔ´Ï´Ù. º¸´Ù ´õ ¾÷±×·¹À̵åµÈ 3¼¼´ë ¹öÀü Tet-On 3G Á¦Ç°À» ¸¸³ª¼¼¿ä!
Lenti-X Tet-On 3G Inducible Expression System (Ŭ¸¯)
- ³·Àº basal expression°ú ÃÖ°íÀÇ ¹ßÇö¿ª°¡¸¦ Á¤È®ÇÏ°Ô Á¶Àý
- Lenti-X Packaging Single Shots (VSV-G) Æ÷ÇÔ: °íÈ¿À², ¾ÈÁ¤¼º ³ôÀº ·»Æ¼¹ÙÀÌ·¯½º ÆÐŰ¡°ú »ý»ê
- Lenti-X Tet-On Advanced¿Í Lenti-X Tet-Off Advanced´Â ´Ù¾çÇÑ cell lines, ºñºÐ¿¼¼Æ÷, Áٱ⼼Æ÷ ¹× Á¶Á÷¿¡ inducible gene expression systemÀ» ±¸Ãà
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DoxycyclineÀ¸·Î Á¶ÀýµÇ´Â ¡±transactivators¡±(Tet-On°ú Tet-Off Advanced)´Â 3 °³ÀÇ ÀÛÀº HSV VP16 ´Ü¹éÁúÀÇ minimal transcription activation domain°ú TetR-À¯·¡ÀÇ DNA-binding domainÀÌ ¿¬°áµÈ À¶ÇÕ ´Ü¹éÁúÀÌ´Ù.
Transactivators´Â ³ôÀº ƯÀ̼ºÀ¸·Î Tetracycline Response Element (TRE)¿Í °áÇÕÇÑ´Ù. TRE´Â PLVX-Tight-PuroÀÇ PTight promoter¿¡ À§Ä¡ÇÏ°í ÀÖÀ¸¸ç ½ÇÇèÀÚÀÇ ¸ñÀû À¯ÀüÀÚ¸¦ Æ÷ÇÔÇÑ´Ù. ½Ã½ºÅÛÀÌ À¯µµµÇ¸é PTight ·ÎºÎÅÍ ¸ñÀû À¯ÀüÀÚ°¡ ³ôÀº ¼öÁØÀ¸·Î Àü»çµÈ´Ù.
Tet-Off Advanced SystemÀº Dox°¡ ÀÖÀ» ¶§ basal state°¡ À¯ÁöµÇ°í, Dox°¡ Á¦°ÅµÇ¸é Àü»ç°¡ ÁøÇà µÈ´Ù. ¹Ý´ë·Î Tet-On Advanced SystemÀº Dox°¡ ÀÖÀ» ¶§ Àü»ç°¡ È°¼ºÈµÈ´Ù (Figure 1).

Figure 1. The Lenti-X Tet-Advanced Systems are highly inducible. Using equal amounts of high-titer supernatants, HeLa cells cultured at the indicated concentrations of Dox were cotransduced for 8 hr with an LVXTight-Puro-Luc lentivirus and an LVX-Tet-On Advanced lentivirus. Cultures were harvested after 48 hr and assayed for luciferase activity. Luciferase was expressed at very high levels, while the basal/uninduced expression level was very low.
Components & Storage Conditions
°¢ Á¦Ç° ±¸¼º¹°°ú º¸°üÁ¶°ÇÀº Certificate of Analysis ¸¦ ÂüÁ¶ÇϽʽÿÀ.