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Inducible System ¡¤ Yeast hybrid
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Matchmaker °ü·ÃÁ¦Ç°
> Aureobasidin A
Aureobasidin A
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Clontech
630466
Aureobasidin A
1 mg
327,000¿ø
Clontech
630499
Aureobasidin A
10 mg
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³»¼ºÀÌ ¾ø´Â ¼¼Æ÷¸¦ Á×ÀÌ´Â °·ÂÇÑ yeast Ç×»ýÁ¦
Aureobasidin A ³»¼ºÀº yeast one- & two-hybrid ¿¬±¸¿¡ ÀÌ»óÀûÀÎ selectable markerÀ̸ç reporterÀÌ´Ù.
Aureobasidin A (AbA)´Â ³·Àº ³óµµ (0.1~0.5 ¥ìg/ml)¿¡¼µµ yeast¿¡ Ä¡¸íÀûÀÎ µ¶¼ºÀ» °¡Áö´Â cyclic depsipeptide°èÀÇ Ç×»ýÁ¦ (Figure 1)ÀÌ´Ù. °¨¼ö¼º ±ÕÁ¾À¸·Î´Â Saccharomyces cerevisiae, Schizosaccharomyces pombe, Candida glabrata, Aspergillus nidulans, A. niger °¡ ÀÖ´Ù. AbA´Â AUR1 À¯ÀüÀÚ¿¡ ÀÇÇØ ¹ßÇöµÇ´Â yeast È¿¼ÒÀÎ inositol phosphorylceramide (IPC) synthase¸¦ ¾ïÁ¦ÇÑ´Ù (1, 2). ÇüÁúÀüȯµÈ yeast¿¡¼ µ¹¿¬º¯ÀÌ À¯ÀüÀÚÀÎ AUR1-CÀÌ ¹ßÇöµÇ¸é yeast°¡ ¾àÁ¦ ³»¼ºÀ» ³ªÅ¸³»µµ·Ï ÇÏ¿© Matchmaker Gold SystemsÀÇ reporter·Î ÀÛ¿ëÇÑ´Ù.
Yeast One- & Two-Hybrid ¿¬±¸¸¦ À§ÇÑ ¿Ïº®ÇÑ Reporter
AbA´Â ³»¼ºÀÌ ¾ø´Â yeast¸¦ ´Ü¼øÈ÷ »ýÀå ¾ïÁ¦½ÃÅ°±â º¸´Ù´Â Á×À̱⠶§¹®¿¡ AbA selectionÀº »ç½Ç»ó nutritional marker (¿¹ : HIS3) ´Üµ¶À¸·Î »ç¿ëÇÏ¿´À» °æ¿ì ¾ö°ÝÇÏÁö ¾ÊÀº screening °úÁ¤À¸·Î ÀÎÇÏ¿© ¹ß»ýµÇ´Â background colony¸¦ Á¦°Å½ÃÄÑ ÁØ´Ù. AbA ±â¹ÝÀÇ selectionÀº ÁøÂ¥ positive cloneÀÇ ¼±º°°ú È®Àο¡ Å©°Ô µµ¿òÀÌ µÈ´Ù. AbA selection ´Üµ¶À¸·Î »ç¿ëÇÏ¿© ¾ö°ÝÇÏÁö ¾ÊÀº ÀÏÂ÷ screening °úÁ¤¿¡¼ ¼±º°µÈ cloneÀ» °Ë»çÇÑ °á°ú, 4 °³ÀÇ ¸ðµç Matchmaker Gold reporter (AUR1-C, HIS3, ADE2, MEL1)¸¦ ¹ßÇöÇÏ´Â ÁøÂ¥ positive°¡ ³ôÀº ºñÀ²·Î È®ÀεǾú´Ù. Á» ´õ ¸¹Àº Á¤º¸´Â Ä«Å»·Î±× Matchmaker Gold Yeast Two-Hybrid System ºÎºÐ¿¡¼ º¼ ¼ö ÀÖ´Ù.
Figure 1. Structure of Aureobasidin A
. Aureobasidin A (MW 1,100) is a cyclic depsipeptide antibiotic isolated from the fungus, Aureobasidium pullulans R106. AbA inhibits the product of the yeast AUR1 gene (inositol phosphorylceramide synthase) and is toxic to S. cerevisiae at low concentrations (0.1 ¥ìg/ml). A dominant mutant allele, AUR1-C, confers resistance to AbA, and its expression can be used as a selectable marker.
Components & Storage Conditions
°¢ Á¦Ç° ±¸¼º¹°°ú º¸°üÁ¶°ÇÀº Certificate of Analysis ¸¦ ÂüÁ¶ÇϽʽÿÀ.
References
1. Takesako, K. et al. (1993) J. Antibiot. (Tokyo) 46(9):1414-20.
2. Hashida-Okado, T. et al. (1996) Mol. Gen. Genet. 251(2):236-244.
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