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Home > ÀüÁ¦Ç°º¸±â > Inducible System ¡¤ Yeast hybrid > Yeast Two Hybrid System (Y2H) > Normalized Y2H Library (Yeast ³»¿¡ µµÀÔµÈ »óÅÂ)

Normalized Y2H Library (Yeast ³»¿¡ µµÀÔµÈ »óÅÂ)

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Clontech
630480
Mate & Plate Library - Universal Human (Normalized)
°ü·ÃÇмú ±¸¸ÅÇϱâ LMOÁ¦Ç° 
1 ml¡¿5
2,580,000¿ø  Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â x33367, x32816
Clontech
630483
Mate & Plate Library - Universal Mouse (Normalized)
°ü·ÃÇмú ±¸¸ÅÇϱâ LMOÁ¦Ç° 
1 ml¡¿5
2,580,000¿ø  Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â x33367, x32816
Clontech
630485
Mate & Plate Library - Universal Drosophila (Normalized)
°ü·ÃÇмú ±¸¸ÅÇϱâ LMOÁ¦Ç° 
1 ml¡¿5
2,580,000¿ø  Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â x33367, x32816
Clontech
630487
Mate & Plate Library - Universal Arabidopsis (Normalized)
°ü·ÃÇмú ±¸¸ÅÇϱâ LMOÁ¦Ç° 
1 ml¡¿5
2,580,000¿ø  Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â x33367, x32816
Clontech
630486
Mate & Plate Library - Human Brain (Normalized)
°ü·ÃÇмú ±¸¸ÅÇϱâ LMOÁ¦Ç° 
1 ml¡¿5
2,580,000¿ø  Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â x33367, x32816

  • Yeast two-hybrid library¸¦ screening Çϱâ À§ÇÑ °¡Àå °£´ÜÇÑ ¹æ¹ý
  • ¸î ÁÖ ³»¿¡ library screening ¿Ï·á - library ÁõÆø°ú transformation ºÒÇÊ¿ä
  • Library nomalization °úÁ¤À» ÅëÇÏ¿© Àú¹ßÇöµÇ´Â cDNAÀÇ ÇÔ·®À» ³ôÀÓ
È¿°úÀûÀÎ »óÈ£ÀÛ¿ë Ž»öÀ» À§Çؼ­´Â Matchmaker Gold Yeast Two-Hybrid System°ú ÇÔ²² Mate & Plate Libraries »ç¿ëÀ» °­·ÂÇÏ°Ô ÃßõÇÑ´Ù (1). ÀÌ Á¦Ç°Àº cDNA library¸¦ ¹Ì¸® yeast¿¡ transformation½ÃÄÑ library ÁõÆøÀ̳ª º°µµÀÇ transformation °úÁ¤¾øÀÌ °¡Àå °£´ÜÇÑ ¹æ¹ýÀ¸·Î two-hybrid screeningÀ» ¼öÇàÇÒ ¼ö ÀÖ´Ù. Mate & Plate library´Â cDNA library·ÎºÎÅÍ prey ´Ü¹éÁúÀ» ¹ßÇöÇÏ´Â Y187 MAT¥ástrain°ú bait ´Ü¹éÁúÀ» ¹ßÇöÇÏ´Â Y2HGold (MATa) reporter strainÀ» mating Çϸé ÇϳªÀÇ yeast¿¡¼­ µÎ °³ÀÇ ´Ü¹éÁúÀ» ¹ßÇö½Ãų ¼ö ÀÖ´Ù. µû¶ó¼­ library¸¦ °¡Áö°í ÀÖ´Â strain°ú bait ´Ü¹éÁúÀ» ¹ßÇöÇÏ´Â strainÀ» ÇϳªÀÇ tube¿¡ ³Ö°í ¹è¾çÇÑ ÈÄ ¼±ÅùèÁö¿¡ µµ¸»Çϸé screening °úÁ¤À» ½ÃÀÛÇÒ ¼ö ÀÖ´Ù (Figure 1).
Normalized & Universal Libraries
Library normalizationÀº ¼¼Æ÷¿¡¼­ »ó´ëÀûÀ¸·Î ´Ù·® ¹ßÇöµÇ´Â cDNA¸¦ ¼±ÅÃÀûÀ¸·Î Á¦°ÅÇÏ¿© ³·Àº ¼öÁØÀ¸·Î ¹ßÇöµÈ cDNAÀÇ »ó´ëÀû ÇÔ·®À» ³ô¿© ÁØ´Ù. Áï, housekeeping gene cDNA µî°ú °°ÀÌ »ó´ëÀûÀ¸·Î dzºÎÇÏ°Ô Á¸ÀçÇÏ´Â cDNA¸¦ °¨¼Ò½ÃÄÑ ¼±º° °úÁ¤¿¡¼­ »ý¼ºµÇ´Â false positive¸¦ Å©°Ô °¨¼Ò½ÃŲ´Ù (1). µû¶ó¼­ ¼±º°À» À§ÇÑ ´õ¿í ´Ù¾çÇÑ ¼öÀÇ cloneÀ» °£ÆíÇÏ°Ô È®º¸ÇÒ ¼ö ÀÖÀ¸¸ç, ¹ßÇö ¼öÁØÀÌ ³·Àº ´Ü¹éÁú°úÀÇ »óÈ£ÀÛ¿ëÀ» °ËÃâÇÒ °¡´É¼ºµµ ³ô¾ÆÁø´Ù.

Universal Library´Â °¡Àå ±¤¹üÀ§ÇÑ À¯ÀüÀÚ ¹ßÇöÀ» ´ëº¯ÇÒ ¼ö ÀÖµµ·Ï Àüü Á¶Á÷ (whole tissues)À¸·ÎºÎÅÍ Á¦ÀÛÀÌ µÇ¾úÀ¸¸ç, ¿ª½Ã normalization µÇ¾î ÀÖ´Ù(2). µû¶ó¼­ ¸ñÀû ´Ü¹éÁú°ú »óÈ£ÀÛ¿ëÇÏ´Â ¹ÌÁöÀÇ ´Ü¹éÁúÀ» ¹ß°ßÇÒ °¡´É¼ºÀÌ ´õ¿í Çâ»óµÇ¾ú´Ù.


Figure 1. Mating and plating. Grow a 50 ml culture of the Y2HGold yeast strain transformed with your bait vector, add a vial of Mate & Plate Library, culture overnight, and plate on medium to select for positive interactions (SD/-Leu/-Trp + AbA + X-¥á-Gal)
Components & Storage Conditions
°¢ Á¦Ç° ±¸¼º¹°°ú º¸°üÁ¶°ÇÀº Certificate of Analysis ¸¦ ÂüÁ¶ÇϽʽÿÀ.
References
1. Mate & Plate Yeast Two-Hybrid cDNA Libraries. (January 2009) Clontechniques XXIV(1):6-7.
2. Shagin, D. A., et al. (2002) Genome Res. 12(12):1953-1942.

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