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Yeast One-Hybrid (Y1H) System

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Á¦Á¶»ç Á¦Ç°ÄÚµå Á¦Ç°¸í ¿ë·® °¡°Ý
(ºÎ°¡¼¼º°µµ)
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Clontech
630491
Matchmaker Gold Yeast One-Hybrid Library Screening System
°ü·ÃÇмú ±¸¸ÅÇϱâ LMOÁ¦Ç° 
1 Set
(Package)
3,639,000¿ø  Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â x33226, x33228, x33365, x33366, x32815, x33213
Clontech
630496
Matchmaker Insert Check PCR Mix 1
°ü·ÃÇмú ±¸¸ÅÇϱâ
100 ȸ
352,000¿ø  Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â x32817
Clontech
630497
Matchmaker Insert Check PCR Mix 2
°ü·ÃÇмú ±¸¸ÅÇϱâ
100 ȸ
352,000¿ø  Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â x32817

  • °¡Àå °íÈ¿À²ÀûÀÎ yeast one-hybrid ½Ã½ºÅÛ
  • »õ·Î¿î Protein-DNA °£ÀÇ »óÈ£ÀÛ¿ë È®ÀÎ
  • Aureobasidin A screening °úÁ¤ Ãß°¡·Î background °¨¼Ò
  • Yeast¿¡¼­ Á÷Á¢ SMART cDNA library Á¦ÀÛ ¹× screening °¡´É
Aureobasidin A·Î ³·Àº background ½ÇÇö
Matchmaker Gold Yeast One-Hybrid Library Screening SystemÀº ¹ÌÁöÀÇ ´Ü¹éÁú°ú DNA °£ÀÇ »óÈ£ÀÛ¿ëÀ» È®ÀÎÇÏ°í ±× ±â´ÉÀ» ÆľÇÇÒ ¼ö ÀÖ´Â °£´ÜÇÏ°í È¿À²ÀûÀÎ ¹æ¹ýÀ» Á¦°øÇÑ´Ù (Figure 1). ¸ðµç Matchmaker Gold SystemÀº Á¤È®ÇÏ°í »ç¿ëÇϱâ Æí¸®ÇÑ reporter·Î Aureobasidin A ³»¼º (AbAr)À» »ç¿ëÇÑ´Ù. ÀÌ »õ·Î¿î reporter´Â Ç×»ýÁ¦ÀÎ Aureobasidin A (AbA)·Î ÀÎÇØ AbA ³»¼ºÀ¯ÀüÀÚ¸¦ ¹ßÇöÇÏÁö ¸øÇÏ´Â yeast¸¦ È¿°úÀûÀ¸·Î Á¦°ÅÇϱ⠶§¹®¿¡ ¸Å¿ì ³·Àº screening background¸¦ °¡´ÉÇÏ°Ô ÇÏ¿´´Ù.
Matchmaker Gold One-Hybrid System
Matchmaker Gold Yeast One-Hybrid system¿¡¼­´Â ¸ñÀû DNA (bait) ¼­¿­À» ¸î ¹ø ¹Ýº¹ÇÏ¿© pAbAi reporter vector¿¡ cloning ÇÑ ÈÄ, bait-specific reporter¸¦ °¡Áø yeast strainÀ» ¸¸µé±â À§ÇÏ¿© pAbAi vector¸¦ homologous recombinationÀ¸·Î Y1HGold yeast genomeÀ¸·Î »ðÀÔ½ÃÄÑ library screeningÀ» À§ÇÑ ¼÷ÁÖ·Î »ç¿ëÇÑ´Ù.


Figure 1. Screening for protein-DNA interactions with the Matchmaker Gold One-Hybrid System. One to three copies of the DNA target sequence are cloned into the pAbAi reporter vector, which is then integrated into the Y1HGold genome to create a bait-specific reporter strain. Activation of the AbA resistance gene (AbAr) occurs if a prey protein from the library binds to the bait sequence.
One-step Library Á¦ÀÛ°ú Screening
Yeast GAL4-transcription activation domain (GAL4 AD prey protein)°ú À¶ÇÕµÇ¾î ¹ßÇöµÇ´Â DNA-binding proteinÀ» Æ÷ÇÔÇÏ´Â cDNA library´Â SMART ±â¼ú°ú homologous recombinationÀ» ÀÌ¿ëÇÏ¿© Y1HGold [Bait-AbAi] reporter strain¿¡¼­ Á÷Á¢ Á¦ÀÛÇÒ ¼ö ÀÖ´Ù. (Figure 2). Prey ´Ü¹éÁúÀÌ bait sequence¿Í °áÇÕÇϸé GAL4 AD´Â AbA ÀúÇ×¼º À¯ÀüÀÚÀÇ ¹ßÇöÀ» È°¼ºÈ­½ÃÄÑ AbA¸¦ Æ÷ÇÔÇÏ´Â ¹èÁö¿¡¼­ yeast°¡ »ýÁ¸ÇÏ°Ô µÈ´Ù. µû¶ó¼­ library screening °úÁ¤¿¡¼­ »ì¾Æ³²Àº yeast cloneÀ¸·ÎºÎÅÍ prey ´Ü¹éÁúÀ» ¹ßÇöÇÏ´Â plasmid¸¦ ½±°Ô ȹµæÇÒ ¼ö ÀÖ°í À̸¦ ÀÌ¿ëÇÏ¿© ´ÙÀ½ °úÁ¤ÀÇ ½ÇÇèÀ» ÁøÇàÇÒ ¼ö ÀÖ°Ô µÈ´Ù.
Colony PCR Kit & Media Set
Colony PCRÀº bait strainÀ» È®ÀÎÇÏ°í screeningÀ» ÅëÇØ È®ÀÎµÈ positive cloneÀ» ºÐ·ùÇϱâ À§ÇÑ °¡Àå ºü¸£°í Æí¸®ÇÑ ¹æ¹ýÀÌ´Ù. Matchmaker Insert Check PCR Mix 1Àº pBait-AbAi ÀçÁ¶ÇÕü°¡ yeast genome¿¡ »ðÀԵǾú´ÂÁö È®ÀÎÇϱâ À§ÇÏ¿© »ç¿ëÇϸç, Matchmaker Insert Check PCR Mix 2´Â onehybrid ¶Ç´Â two-hybrid screening °úÁ¤¿¡¼­ ¾òÀº positive cloneÀ» ºü¸£°Ô ºÐ¼®Çϱâ À§ÇÏ¿© »ç¿ëÇÑ´Ù. Yeast Media Sets 1°ú 1 Plus´Â Matchmaker Gold One-Hybrid ½ÇÇè¿¡ ÇÊ¿äÇÑ ¹èÁö¸¦ pouch ÇüÅ·Π±¸¼ºÇÑ Á¦Ç°ÀÌ´Ù.


Figure 2. Use SMART technology and yeast biology to simultaneously construct and screen your library directly in yeast. SMART technology creates a pool of cDNA harboring ends that are homologous to the linearized prey vector, pGADT7-Rec. The cDNA pool and pGADT7-Rec are cotransfomed into your Y1HGold-Bait reporter strain, and undergo homologous recombination within the yeast. Yeast cells are then plated on SD/-Leu/+AbA to select for colonies that express reporters as a result of positive Y1H interactions.
Components & Storage Conditions
°¢ Á¦Ç° ±¸¼º¹°°ú º¸°üÁ¶°ÇÀº Certificate of Analysis ¸¦ ÂüÁ¶ÇϽʽÿÀ.

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