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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)
1 ml¡¿5
2,064,000¿ø
2,580,000¿ø
11.01 ~ 12.27
,
Clontech
630483
Mate & Plate Library - Universal Mouse (Normalized)
1 ml¡¿5
2,064,000¿ø
2,580,000¿ø
11.01 ~ 12.27
,
Clontech
630485
Mate & Plate Library - Universal Drosophila (Normalized)
1 ml¡¿5
2,064,000¿ø
2,580,000¿ø
11.01 ~ 12.27
,
Clontech
630487
Mate & Plate Library - Universal Arabidopsis (Normalized)
1 ml¡¿5
2,064,000¿ø
2,580,000¿ø
11.01 ~ 12.27
,
Clontech
630486
Mate & Plate Library - Human Brain (Normalized)
1 ml¡¿5
2,064,000¿ø
2,580,000¿ø
11.01 ~ 12.27
,
Product index
Tet À¯µµ ¹ßÇö ½Ã½ºÅÛ
[°³¿ä] Tet-on system
[¼±Åð¡À̵å] Tet Syste..
Tet-On 3G À¯µµ ¹ßÇö ½Ã..
Lenti-X Tet-On 3G Indu..
Tet-On 3G Cell Line
Tet Approved FBS
Tet °ü·Ã Ç×»ýÁ¦
Selection marker (Line..
Tet-One Inducible Expr..
TetR Ç×ü
[Legacy] Tetracycline ..
Tet-Off Cell Lines
pTRE-Tight Vectors
pTet-tTS Vectors
IRES Tet À¯µµ¹ßÇö ½Ã½º..
iDimerize Inducible System..
[°³¿ä] Inducible Prote..
iDimerize Antibodies
Inducible Expression S..
Heterodimer System
Homodimer System
Regulated Transcriptio..
Reverse Dimerization S..
´Ü¹éÁú ¹ßÇö·® Á¶Àý ½Ã½ºÅÛ ..
ProteoTuner System
Tet-ProteoTuner Vector..
[°³¿ä] Yeast Hybrid System
[FAQ] Matchmaker Gold ..
[°³¿ä] Matchmaker Gold..
Yeast Two Hybrid System (Y..
Y2H Library Constructi..
Yeast Culture media (T..
Yeast Two-Hybrid (Y2H)..
Y2H Library (Yeast ³»..
Normalized Y2H Library..
Yeast Two hybrid vecto..
pBridge Vector
Yeast One-Hybrid (Y1H)
Yeast Culture Media (O..
Yeast One-Hybrid (Y1H)..
Matchmaker °ü·ÃÁ¦Ç°
Yeast Culture Media
Yeast Culture Media (..
Aureobasidin A
Yeast Plasmid Isolati..
Matchmaker Ç×ü
Insert Check Kit (prey..
Yeast Transformation M..
X-¥á-Gal
Yeast Transformation S..
Mammalian Cell Assay K..
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.
Keyword :
Yeast Two Hybrid System (Y2H)
Y2H Library Construction System
Yeast Culture media (Two-Hybrid ¿ë)
Yeast Two-Hybrid (Y2H) System
Y2H Library (Yeast ³»¿¡ µµÀÔµÈ »óÅÂ)
Normalized Y2H Library (Yeast ³»¿¡ µµÀÔµÈ »óÅÂ)
Yeast Two hybrid vectors
pBridge Vector