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EST, Microarray, and Plant ScienceÀÌ »ó Çù / ¹Ú»ç »ý¸í°øÇבּ¸¼Ò, ¿¬±¸¿ø Tel. 042-860-4466 Fax. 042-860-4608 E-mail. sanghyeobl@yahoo.co.kr ÃÖ±Ù µé¾î ±Þ¼ÓÈ÷ ¹ß´ÞµÈ DNA sequencing technology¿Í À̸¦ ¹ÙÅÁÀ¸·Î ÇÑ ´ë·®ÀÇ À¯ÀüÀÚ ¼¿ ºÐ¼® ±â¹ýÀÇ È®¸³Àº ±âÁ¸ÀÇ ºÐÀÚ »ý¹°ÇÐÀû ½ÇÇè¿¡ Áö´ëÇÑ ¿µÇâÀ» ¹ÌÃÆ´Ù. µû¶ó¼ geneticÀ̳ª reverse-geneticÀ» ÀÌ¿ëÇؼ Çѵΰ³ÀÇ À¯ÀüÀÚÀÇ ±â´ÉÀ» ¹àÈ÷´Â ±âÁ¸ÀÇ ¹æ½ÄÀ¸·Î´Â ´ë·®À¸·Î ½ñ¾ÆÁö´Â À¯ÀüÀÚ Á¤º¸¸¦ ó¸®ÇÒ ¼ö ¾ø´Â ÇÑ°è »óȲ¿¡ óÇß´Ù. µû¶ó¼ À̸¦ ±Øº¹ÇÒ »õ·Î¿î ¹æ¹ýµéÀÌ °³¹ßµÇ°í ÀÖ°í ±× Áß °¡Àå °·ÂÇÑ ¹æ¹ýÀÌ microarray technologyÀÌ´Ù. Genomics ÇöÀç µÎ Á¾ÀÇ ½Ä¹°Ã¼, ¾Ö±âÀå´ë (Arabidopsis thaliana), º (Oryza sativa)¿¡¼ Àüü ¿°±â¼¿ÀÌ ¹àÇôÁ³´Ù (Arabidopsis Genome Initiative, 2000). ÀÌ·¯ÇÑ Àüü Áö³ðÀÇ ¿°±â¼¿ È®º¸´Â ½Ä¹°Ã¼ ÇÑ °³Ã¼¿¡ ´ëÇÑ systemicÇÑ ºÐ¼®À» °¡´ÉÄÉ ÇØ ÁÖ¸®¶ó ¿¹ÃøµÈ´Ù. Áï È®º¸µÈ ¿°±â¼¿À» ¹ÙÅÁÀ¸·Î À¯ÀüÀÚ ºÎºÐÀ» ¿¹ÃøÇØ ³¾ ¼ö ÀÖÀ¸¸ç Àüü À¯ÀüÀÚ¿¡ ´ëÇÑ ±â´É ºÐ¼® ¹× À̵é À¯ÀüÀÚµéÀÇ »óÈ£ °ü°è ºÐ¼®À» ÅëÇØ »ý¸íÇö»ó¿¡ ´ëÇÑ ÀÌÇØÀÇ ÆøÀ» ³ÐÈú °ÍÀÌ´Ù. ±×·¯³ª ¿ì¸®°¡ ¿¬±¸ ´ë»óÀ¸·Î ÇÏ´Â ¸ðµç ½Ä¹°Ã¼¸¦ ´ë»óÀ¸·Î Àüü ¿°±â¼¿À» ½ÃÇàÇϱ⿡´Â »ó´çÇÑ ºñ¿ëÀÌ ÇÊ¿äÇÏ´Ù. µû¶ó¼ ´ë¾ÈÀ¸·Î ºÎ»óÇÏ´Â °ÍÀÌ EST (Expressed Sequence Tag)ÀÌ´Ù. EST¶õ ¸» ±×´ë·Î ¹ßÇöµÇ´Â À¯ÀüÀÚ ´ÜÆíÀÌ´Ù. ÀÌ·¯ÇÑ EST´Â ƯÁ¤ Á¶Á÷À̳ª ȯ°æ Á¶°Ç¿¡¼ ¹ßÇöµÇ´Â mRNA¸¦ ¿ªÀü»ç (Reverse Transcription) ¹ÝÀÀÀ» ÅëÇØ »ý¼ºµÈ cDNA¸¦ 5′- À̳ª 3′- end ºÎºÐÀ» ¿°±â¼¿ÇÔÀ¸·Î½á »ý¼ºµÈ´Ù (Boguski and Schuler, 1995). ±×·¯¹Ç·Î EST´Â »ùÇøµÇÒ ¶§¿¡ ¹ßÇöµÇ´Â À¯ÀüÀÚ¿¡ ´ëÇÑ snapshot¸¸ Á¦°øÇÒ »ÓÀÌ°í, intronÀ̳ª regulatory DNA¿¡ ´ëÇÑ Á¤º¸´Â Á¦°øÇÏÁö ¸øÇÑ´Ù (Lee, S. P. and Lee, K. H. 2000). ±×·³¿¡µµ ºÒ±¸ÇÏ°í È®º¸ÀÇ ¿ëÀ̼º ¶§¹®¿¡ cDNA libraryÀÇ Áõ°¡¿Í ´õºÒ¾î ESTÀÇ ¼ö°¡ ±âÇϱ޼öÀûÀ¸·Î ´Ã°í ÀÖ´Ù. 2000³â 12¿ùÀ» ±âÁØÀ¸·Î dbEST (http://www. ncbi.nlm.nih.gov/ dbEST/)¸¦ º¸¸é 6,200,000°³ ÀÌ»óÀÇ ÀÇ EST°¡ ÀúÀåµÇ¾î ÀÖ´Ù. ÀÌ Áß ½Ä¹°Ã¼°¡ 102Á¾ÀÌ¸ç ´ëÇ¥ÀûÀÎ Á¾Àº Glycine max, Arabidopsis thaliana, Lycopersicon esculentum, Zea mays, Oryza sativa, Sorghum bicolor, Triticum aestivum, Solanum tuberosum µîÀÌ´Ù (Quackenbush, J. et al., 2001). ¾Ö±âÀå´ëÀÇ °æ¿ì¸¦ º¸¸é, 105,733°³ÀÇ redundancy¸¦ Æ÷ÇÔÇÑ EST°¡ Àüü 25,498°³ÀÇ À¯ÀüÀÚ Áß 60%¸¦ ³ªÅ¸³»´Â °ÍÀ¸·Î ¹àÇôÁ³´Ù (Arabidopsis Genome Initiative, 2000). ÇöÀç ±¹³»¿¡¼µµ ¹èÃß, Àλï, °íÃ߸¦ Áß½ÉÀ¸·Î EST¸¦ È®º¸ÇÏ´Â ÀÛ¾÷ÀÌ È°¹ßÈ÷ ÁøÇà ÁßÀÌ´Ù. Functional genomics ´ë·®À¸·Î ½ñ¾ÆÁö´Â À¯ÀüÀÚÀÇ ±â´ÉÀ» ±âÁ¸ÀÇ ÇüÁúÀüȯü ¹æ¹ýÀ» ÀÌ¿ëÇؼ ¹àÈ÷´Â °ÍÀÌ °ÅÀÇ ºÒ°¡´ÉÇØÁ³´Ù. µû¶ó¼ ´ë·®À¸·Î À¯ÀüÀÚ ±â´ÉÀ» ¹àÈ÷´Â ÀÛ¾÷ÀÌ tagging lineÀ» ÀÌ¿ëÇؼ ½ÃµµµÇ¾îÁ® ¿ÔÀ¸³ª ½Ä¹°Ã¼ ÀÚü°¡ °®°í ÀÖ´Â À¯ÀüÀÚÀÇ ±â´ÉÀû Áߺ¹ (functional redundancy)¼º ¶§¹®¿¡ Å©°Ô ¼º°øÀ» °ÅµÎÁö ¸øÇÏ´Â ½ÇÁ¤ÀÌ´Ù. µû¶ó¼ ÀÌ¿¡ ´ëÇÑ ´ë¾ÈÀ¸·Î ¹ÙÀÌ·¯½º¸¦ ÀÌ¿ëÇÑ gene silencing (Virus Induced Gene Silencing)À» ÀÌ¿ëÇÏ´Â °ÍÀÌ ½ÃµµµÇ°í ÀÖ´Ù (Dalmay et al., 2000). ÀÌ ¹æ¹ýÀÇ ÀåÁ¡Àº sense ¿Í antisense »ó°ü¾øÀÌ 80% ÀÌ»óÀÇ sequence identity¸¦ °¡Áö¸é gene family memberÀÏÁö¶óµµ µ¿½Ã¿¡ °¢ family member À¯ÀüÀÚÀÇ ¹ßÇöÀ» ¾ïÁ¦ÇÑ´Ù. µû¶ó¼ ½Ä¹°Ã¼Ã³·³ ´Ù·®ÀÇ gene family°¡ Á¸ÀçÇÏ´Â system¿¡¼´Â ¾ÆÁÖ À¯¿ëÇÑ ¹æ¹ýÀÌ´Ù. ±×·¯³ª ÀÌ·± ÀÌÁ¡¿¡µµ ºÒ±¸ÇÏ°í ´ã¹è (Nicotiana bentamia ¿¡¼¸¸ È¿À²ÀûÀ¸·Î ÀÛ¿ë) ¿Ü¿¡´Â ¾ÆÁ÷ È¿°úÀûÀ¸·Î ÀÌ¿ëµÇÁö´Â ¸øÇÏ°í ÀÖ´Ù. µû¶ó¼ ´ë¾ÈÀ¸·Î À¯ÀüÀÚ ¹ßÇö¾ç»ó ºÐ¼® (gene expression pofiling), ¾Æ¹Ì³ë»ê ¼¿ »óµ¿¼º ºÐ¼®(sequence analysis), ºñ±³ À¯Àüü ºÐ¼® (comparative genomics or synteny) µîÀÇ »ó´çÈ÷ ´Ù¾çÇÑ computationÇÑ ¹æ¹ýÀ» ÀÌ¿ëÇؼ ±â´ÉÀ» À¯ÃßÇÏ´Â ½Ãµµ°¡ ÁøÇàµÇ°í ÀÖ´Ù. À¯ÀüÀÚ ¹ßÇö¾ç»ó ºÐ¼® mRNA ¼öÁØ¿¡¼ÀÇ À¯ÀüÀÚ ¹ßÇöÀ» ºÐ¼®ÇÏ´Â ¹æ¹ýÀº ¿©·¯ °¡Áö°¡ ÀÖÀ¸¸ç Å©°Ô Northen-blot (Alwine et al., 1977), differential display (Liang and Pardee, 1992), dotblot analysis (Lennon and Lehrach, 1991), Serial analysis of gene expression (SAGE; Velculescu et al., 1995) µîÀ¸·Î ³ª´©¾îÁø´Ù (Table 1). À§¿¡¼ ¾ð±ÞÇÑ ±âÁ¸ÀÇ ºÐ¼®¹ýÀ¸·Î´Â ´ë·®ÀÇ À¯ÀüÀÚ¸¦ µ¿½Ã¿¡ ºÐ¼®Çϱ⿡ ±â¼úÀûÀÎ ÇÑ°è°¡ ÀÖ¾ú´Ù. µû¶ó¼ À̸¦ °¡´ÉÄÉÇÒ ¹æ¹ýÀ¸·Î DNA microarray technology°¡ »õ·Ó°Ô ´ëµÎµÇ¾ú´Ù (Schena et al., 1995, Watson et al., 1998, Duggan et al., 1999). Microarray¸¦ immobolized target sequence¿¡ µû¶ó DNA fragement based microarray (Schena et al., 1995) ¿Í oligonucleotide based microarray (Lipshutz et al., 1999)·Î Å©°Ô ³ª´©¾î º¼ ¼ö ÀÖ´Ù. Oligonucleotide based micrarrayer´Â ¾ÆÁÖ ÂªÀº oligomer (20-25mer)¸¦ »ç¿ëÇϹǷΠsingle nucleotide mismatch¿¡ ±²ÀåÈ÷ sensitiveÇϹǷΠgenotyping µî¿¡ ÀûÇÕÇÏ´Ù. ±×¸®°í oligo chipÀ» Á¦ÀÛÇϱâ À§Çؼ´Â Á¤È®ÇÑ sequenec data°¡ ÇʼöÀûÀÌ´Ù. ÀÌ¿¡ ¹ÝÇØ DNA fragment based microarray´Â sequence informationÀÌ ÇʼöÀûÀÌÁø ¾Ê´Ù. µû¶ó¼ Àü¹ÝÀûÀ¸·Î sequence informationÀÌ ºÎÁ·ÇÑ ½Ä¹°Ã¼¸¦ ´ë»óÀ¸·Î À¯ÀüÀÚ ¹ßÇöÀ» ºÐ¼®ÇÒ ¶§¿¡´Â DNA chipÀÌ À¯¸®ÇÏ´Ù. µû¶ó¼ ¿©±â¿¡¼´Â ÁÖ·Î cDNA microarray¿¡ ´ëÇØ ¾ð±ÞÇÒ °ÍÀÌ´Ù. Microarray¿¡ ´ëÇÑ ±â¼úÀûÀÎ Ãø¸é (Á¦ÀÛ¹æ¹ý°ú »ç¿ë¹ý), commercial supplier, instruments¿¡ ´ëÇÑ ÀÚ¼¼ÇÑ ³»¿ëÀº ´ÙÀ½ ³í¹®À» ÂüÁ¶Çϸé À¯¿ëÇÑ Á¤º¸¸¦ ¾òÀ» ¼ö ÀÖ´Ù (Lemieux et al., 1998; Eisen and Brown 1999; Marshall 1998). DNA microarray Microarray´Â À¯¸® slided À§¿¡ DNA Á¶°¢À» ±â°è¸¦ ÀÌ¿ëÇؼ ¾ÆÁÖ Á¶¹ÐÇÏ°Ô spottingÇÔÀ¸·Î½á Á¦À۵ǾîÁø´Ù (Fig. 1). DNA Á¶°¢ÀÇ source´Â ´ë°³ genome sequencing °á°ú·Î ³ª¿Â EST clones, anonymous cDNA clones, genomic clones, DNA amplified from open reading frames (ORF) µîÀε¥ °¢±â ÇÊ¿ä·Î ÇÏ´Â ½ÇÇè ¸ñÀû¿¡ µû¶ó Á¤ÇØÁø´Ù. ÇöÀçÀÇ ±â¼úÀûÀÎ ¼öÁØÀ¸·Î´Â 10,000 spots/3.24 cm2±îÁö spottingÀÌ °¡´ÉÇÔÀ¸·Î º¸Åë glass slide (1X3 inch)¿¡ 20,000~25,000 °³ À¯ÀüÀÚ¸¦ ½ÉÀ» ¼ö ÀÖ´Ù (Kehoe et al., 1999). Microarray´Â Á¦ÀÛ ½Ã spottingÇÏ´Â ¹æ¹ý¿¡ µû¶ó Åë»óÀûÀ¸·Î Pin?¹æ½Ä°ú Ink-jet?¹æ½ÄÀ¸·Î ³ª´¶´Ù. Pin?¹æ½ÄÀº multiple pin¿¡ DNA¸¦ ¹¯Çô¼ solidÇÑ slide Ç¥¸é¿¡ Á÷Á¢ÀûÀ¸·Î spottingÇÏ´Â ¹æ¹ýÀÌ°í, Ink-jet?¹æ½ÄÀº pizoelectricÀ̳ª solenoid valve¸¦ ÀÌ¿ëÇØ ºÐ»çÇؼ slide Ç¥¸é¿¡ DNA¸¦ ºÎÂø½ÃŲ´Ù (van Hal et al., 2000). ÇöÀç \\Ink-jet\\¹æ½ÄÀÇ ±â¼úÀûÀÎ ÇÑ°è·Î \\Pin\\¹æ½ÄÀÌ ÁÖ·Î »ç¿ëµÇ¾îÁö°í ÀÖÀ¸¸ç À̸¦ ÀÀ¿ëÇÑ Affemetrix»çÀÇ \\Pin and ring\\¹æ½ÄÀº ¿ì¼öÇÑ spot ÀçÇö¼ºÀ» º¸¿©ÁÖ°í ÀÖ´Ù. Microarra Á¦ÀÛ ½Ã DNA°¡ Á÷Á¢ ºÎÂøµÇ¾îÁö´Â glass slide Ç¥¸éÀº DNA¿ÍÀÇ °áÇÕÀ» ½±°ÔÇϱâ À§Çؼ ÈÇÐÀûÀ¸·Î 󸮵ǾîÁö´Âµ¥ ÁÖ·Î poly-L-lysine, amine, aldhyde µîÀÌ ÀÌ¿ëµÈ´Ù (Schena et al., 1995; Schena et al., 1996; Rogers et al., 1999). ÀÌ·¸°Ô positively ó¸®µÈ glassÀÇ Ç¥¸é°ú negatively charged DNA Á¶°¢Àº ½±°Ô °áÇÕÀ» Çü¼ºÇÑ´Ù. ÀÌ¿Ü¿¡µµ slided Ç¥¸éÀ» polyacrylamide gel pad, nylon membrane, nitrocellulose, other polymer¸¦ ÀÌ¿ëÇØ coating ÇÏ·Á´Â ³ë·ÂÀÌ ÇàÇØÁö°í ÀÖ´Ù (Prounikov et al., 1998). ÀÌ·¸°Ô Á¦ÀÛµÈ microarray´Â Çü±¤ dye (ÁÖ·Î Cy3¿Í Cy5 ÀÌ¿ë)·Î ¿°»öµÈ mRMA À¯·¡ÀÇ probeÀ¸·Î hybridizationµÈ´Ù (Fig 1). ÀÌÈÄ microarray¸¦ washing ÇÑ ÈÄ scanner¿¡¼ °¢±â ÀûÇÕÇÑ ÆÄÀåÀÇ laser¿¡¼ ¹ß±¤À» ½ÃŲ ÈÄ °¢ spot¿¡¼ ³ª¿À´Â Çü±¤ÀÇ ¼¼±â¸¦ Æ÷ÂøÇÑ´Ù (Fig. 1). ¿©±â¿¡¼ detect µÇ´Â signalÀÇ ¼¼±â´Â °¢ spotÀÇ ½ÇÁ¦ÀûÀÎ mRNAÀÇ ¹ßÇö·®°ú Á¤·®ÀûÀ¸·Î ºñ·Ê¸¦ ÇÑ´Ù. ÀÌ·¯ÇÑ Çü±¤ dye °¨Áö¹ýÀº °¨µµ°¡ ¶Ù¾î³ª ±â¼úÀûÀ¸·Î 500,000 ÁßÀÇ ÇÑ °³ÀÇ mRNA transcript±îÁö °¨ÁöÇÒ ¼ö ÀÖ´Ù°í ÇÑ´Ù (Schena et al., 1996). ¶ÇÇÑ ´Ù¸¥ excitation°ú emission ÆÄÀåÀ» °¡Áø Çü±¤ dye¸¦ »ç¿ëÇÔÀ¸·Î µÎ sample (»ý¸®ÀûÀ̳ª À¯ÀüÀûÀ¸·Î ´Ù¸¥ mRNA populations) °£ÀÇ ºñ±³°¡ ¿ëÀÌÇÏ´Ù. ½ÇÁ¦ÀûÀ¸·Î microarray ½ÇÇè ½Ã µÎ °³ÀÇ ´Ù¸¥ dye·Î labelingµÈ probe¸¦ ÇÔ²² ¼¯¾î¼ ÇϳªÀÇ microarray¿¡´Ù hybridizationÀ» ÇÔÀ¸·Î hybridization °úÁ¤¿¡¼ ¹ß»ýÇÒ ¼ö ÀÖ´Â ½ÇÇèÀûÀÎ ¿ÀÂ÷¸¦ ÁÙÀÏ ¼ö ÀÖ´Â ÀÌÁ¡ÀÌ ÀÖ´Ù (Fig 1). Çü±¤ dyeÀÇ ¿©·¯ ÀåÁ¡¿¡µµ ºÒ±¸ÇÏ°í photobleachingÀ̶ó´Â ´ÜÁ¡ÀÌ ¿©·¯ °¡Áö ¹®Á¦Á¡À» ¾ß±âÇؼ dataÀÇ ºÐ¼®À» ¾î·Æ°Ô ÇÏ°í ÀÖ´Ù. ½Ç·Ê·Î Fig. 2¿¡¼ º¸µíÀÌ Cy5 dye°¡ Cy3 dyeº¸´Ù Àü¹ÝÀûÀ¸·Î °ÇÑ background intensity¸¦ º¸ÀÌ°í photobleaching¿¡ ´õ ¹Î°¨ÇÏ°Ô ¹ÝÀÀÇÑ´Ù. µû¶ó¼ ½ÇÁ¦·Î data¸¦ ºÐ¼®ÇÒ ¶§´Â ÀÌ·± Çö»óÀ» °¨¾ÈÇÏ¿© data¸¦ º¸Á¤ÇØ ÁÖ¾î¾ß Çϴµ¥ À̸¦ normalizationÀ̶ó°í ÇÏ¸ç ¿©·¯ °¡Áö ¹æ¹ýÀ» ÅëÇØ ÀÌ·ç¾îÁø´Ù (Alon et al., 1999; Eisen et al., 1998; Schena et al., 1996). ½ÇÇèÀûÀ¸·Î´Â dye swapping À̶ó°í Çؼ µ¿ÀÏÇÑ ½ÇÇèÀ» µÎ ¹ø ¹Ýº¹Çϴµ¥ ÀÌ ¶§ °¢ Çü±¤ dye¸¦ ¼·Î ¹Ù²Ù¾î¼ ½ÇÇèÇؼ ³ª¿Â µ¥ÀÌÅ͸¦ ºñ±³ÇÔÀ¸·Î½á dataÀÇ quality¸¦ ³ôÀÌ´Â ¹æ¹ýÀÌ ½ÃÇàµÇ°í ÀÖ´Ù. Characteristics of mcroarray data sets Microarray dataÀÇ ÇÑ °¡Áö ƯÀÌÁ¡Àº ¸¹Àº µ¿ÀÏÇÑ microarray·ÎºÎÅÍ ¾ò¾îÁö±â ¶§¹®¿¡ ¿©·¯ ½ÇÇèÀÚ °£¿¡ µ¥ÀÌÅ͸¦ °øÀ¯ÇÒ ¼ö ÀÖ´Ù. ±×·¯¹Ç·Î stanford microarray database (SMD;http://afgc.stanford.edu/afgc_html/SMDguide/SMD.html) °°Àº centralized databases¸¦ ¸¸µé¸é ¿©·¯ ½ÇÇèÀÚ°¡ data-miningÀ» ÅëÇØ ÀÚ±âµéÀÌ ¿øÇϴ Ư¡ÀûÀÎ ¹ßÇö ÆÐÅÏÀ» º¸ÀÌ´Â À¯ÀüÀÚ ¹«¸®¸¦ ÃßÃâÇÒ ¼öµµ ÀÖ´Ù. ÀÌó·³ parallel ºÐ¼®ÀÌ °¡´ÉÇÑ microarray data´Â ±²ÀåÈ÷ powerfulÇÑ Á¤º¸¸¦ Á¦°øÇÏÁö¸¸ ÀÌ ¶§¹®¿¡ microarray ½ÇÇè ½Ã ÁÖÀÇÇØ¾ß ÇÒ ¿©·¯ ¿äÀÎ (Fig 3)µéÀÌ Àִµ¥ ±× Áß Áß¿äÇÑ Á¡À» »ìÆ캸¸é ¾Æ·¡¿Í °°´Ù. (1) °¢ ½ºÅܸ¶´Ù quality controlÀÌ ÇÊ¿äÇÏ´Ù. (2) ½ÇÇè ½Ã replicationÀÌ ÇÊ¿äÇÏ´Ù. (3) ÇÑ °³ÀÇ microarray¿¡ duplicateÇؼ DNA¸¦ spotting ÇÏ´Â °ÍÀÌ ¹Ù¶÷Á÷ÇÏ´Ù (4) Ç¥ÁØÈµÈ protocol (Àç·á Áغñ, labeling, hybridization, washing process)À» µû¶ó¼ ÇÑ´Ù. ÀÌ·¸°Ô Ç¥ÁØÈµÈ ½ÇÇèÀ» ÅëÇØ »ý»êµÈ dataµéµµ ¼·Î ºñ±³ ºÐ¼®Çϱâ À§Çؼ´Â Ç¥ÁØÈµÈ ¹æ½ÄÀ» µû¶ó¼ database¸¦ ±¸ÃàÇØ¾ß ÇÑ´Ù. ÇöÀç data Ç¥ÁØÈ ¹æ½ÄÀÌ ÃßÁøÁßÀε¥ Á¶¸¸°£ ¹ßÇ¥µÉ ¿¹Á¤ÀÌ°í, ¾î´À Á¤µµ ÃÖ¼ÒÇÑÀÇ Ç¥ÁØÈ ¹æ¾ÈÀº °ø°³µÇ¾î ÀÖÀ¸¸ç MIAM (Minimum Information About A Microarray Experiment)·Î ºÒ¸°´Ù (http://www. cbil.upenn.edu/Ontology/MIAME_MAML. htm; Kolchanov et al., 1999; Stoeckert et al., 2001). Microarray ½ÇÇèÀº Áö±Ý±îÁö »ý¹°ÇÐÀÚµéÀÌ Á¢Çغ¸Áö ¸øÇÑ ¹æ´ëÇÑ data¸¦ »ý»êÇس»°í ÀÖ°í ÇöÀçÀÇ Åë°èÇÐÀû ºÐ¼® ´É·ÂÀÌ À̸¦ µû¶ó°¡Áö ¸øÇÏ°í ÀÖ¾î¼ bottleneck Çö»óÀ» ¾ß±âÇÏ°í ÀÖ´Ù (Ermolaeva et al., 1998). ÀÌ·± Bottleneck Çö»óÀº DNA sequencing ºÐ¾ß¿¡¼µµ °æÇèÇÑ °ÍÀ¸·Î DNA sequences°¡ database¿¡ ÃàôµÇ±â ½ÃÀÛÇÑ ÈÄ ¾à 15³â ÀÌÈÄ°¡ Áö³ª¼¾ß blast °°Àº ¿°±â ¼¿ ºÐ¼® software°¡ routineÇÏ°Ô »ç¿ëµÇ°í ÀÖ´Ù (Xiang and Chen, 2000). ÀÌ·± °æÇè¿¡ ºñÃß¾î ºñ¾àÀûÀÎ ±â¼úÀÇ ¹ßÀü¿¡µµ ºÒ±¸ÇÏ°í »ó´çÇÑ ½Ã°£ÀÌ Áö³ª¾ß detailÇÐ interactiveÇÑ ºÐ¼®ÀÌ °¡´ÉÇØÁö¸®¶ó ¿©°ÜÁø´Ù. Bioinformatics in cDNA microarray ÇöÀç bioinformatics technologyÀÇ ±Þ°ÝÇÑ ¹ß´Þ¿¡µµ ºÒ±¸ÇÏ°í À¯ÀüÀÚ ¹ßÇöÀÇ parallel ºÐ¼®ÀÌ ½±»ç¸® ÀÌ·ç¾îÁöÁö´Â ¾Ê°í ÀÖ´Ù. ±×·¯³ª Á¦ÇÑµÈ ¹üÀ§¿¡¼ »ó¿ëÈµÈ ºÐ¼® toolÀÌ ¿©·¯ °³ ½ÃÆǵǰí ÀÖÀ¸³ª ÇöÀç ¸¹Àº º¸¿ÏÀÌ ÇÊ¿äÇÏ´Ù. ƯÈ÷ laboratory information management, image analysis, expression pattern recognition, comparison of larger number of different experimental samples, statistical analysis, visualization, construction of database µî¿¡ °üÇؼ ¸¹Àº ¹ßÀüÀÌ ÇÊ¿äÇÏ´Ù (Xiang and Chen., 2000). LIMS (Laboratory Information Management System) ´Â experomental material Á¤º¸ÀÇ È帧À» °ü¸®ÇÏ´Â systemÀ¸·Î microarray¿¡ °üÇÑ Àü¹ÝÀûÀÎ Á¤º¸¸¦ Æ÷ÇÔÇÏ°í ÀÖ´Ù (Ermolaeva et al., 1998). ½Ç·Ê·Î¼ ArrayDB (http://www. nhgri.nih.gov/DIR/LCG/15K/HTML/)¸¦ º¸¸é microarray¿Í cDNA clone (clone ID, title, database hyperlink)¿¡ °üÇÑ Á¤º¸, probe Á¤º¸ (sample source, experiment condition, flour type), image information (raw scanned image, intensity and ratio for each target clone) µîÀ» Æ÷ÇÔÇÏ°í ÀÖ´Ù. Microarra image analysis´Â °¢ cDNA target locationÀ» ÀÎÁöÇÏ°í °¢ probeÀÇ intensity ¿Í ratio¸¦ »êÃâÇؼ clone information°ú link½ÃÄѼ ½ÇÇèÀÚ°¡ ¸ðµç ÅëÇÕµÈ Á¤º¸¸¦ ½±°Ô ¾Ë ¼ö ÀÖ°Ô ÇÑ´Ù. ±×·¯³ª ½ÇÁ¦ÀûÀ¸·Î´Â printingÇÑ pin type, hybridization ¹æ¹ý, analysis softwareÀÇ »óÀ̼º ¶§¹®¿¡ °¢°¢ÀÇ ºÐ¼® ¹æ¹ýÀº ¸Å¿ì ´Ù¸£´Ù (Bowtell, 1999; Chen et al., 1997). Gene expression analysis ºÐ¼®Àº ÇöÀç ¿©·¯ Åë°èÀû ¹æ¹ýÀ» ÀÌ¿ëÇϴµ¥ ´ëÇ¥ÀûÀ¸·Î »ç¿ëµÇ´Â °ÍÀÌ hierarchical clustering, k-mean-based anlysis, self organized map (SOM), principle component analysis (PCA) µîÀÌ´Ù (Eisen et al., 1998; Sherlock 2000). ÇöÀç »ç¿ëµÇ¾îÁö´Â ´ëÇ¥ÀûÀÎ ºÐ¼® ¹æ¹ýÀº clustering analysis¶ó°í Çؼ Co-regulationµÇ´Â À¯ÀüÀÚ¸¦ ¹ß°ßÇÏ´Â µ¥ Ä¡ÁßÇÏ°í ÀÖÀ¸¸ç ÀÌ·± À¯ÀüÀÚ ¹«¸®µéÀº °øÅëÀûÀÎ promoter motif, transcription factor, function, signal transduction pathway¸¦ °¡Áú °¡´É¼ºÀÌ »ó´çÈ÷ ³ô´Ù (DeRisi et al., 1997). Practical approaches of microarray (1) ÇöÀç »ó´çÈ÷ ¸¹Àº sequencing project°¡ ½Ä¹°Ã¼¸¦ Áß½ÉÀ¸·Î ÁøÇàÁßÀ̸ç, ÀÌÁß ¸¹Àº À¯ÀüÀÚ°¡ sequencing homology¸¦ ¹ÙÅÁÀ¸·Î ±â´É (functional annotation)ÀÌ ºÎ¿©µÇ°í ÀÖ´Ù. ±×·¯³ª microarray¸¦ ÀÌ¿ëÇؼ ¿©·¯ °¡Áö ȯ°æ, ¹ß´Þ ´Ü°è¿¡¼ÀÇ À¯ÀüÀÚÀÇ ¹ßÇö ¾ç»óÀ» ºñ±³ ºÐ¼®ÇÏ¸é ´Ü¼øÈ÷ sequence based functional annotation º¸´Ù Á¤È®È÷ ±â´ÉÀ» ¿¹ÃøÇÒ ¼ö ÀÖÀ» °ÍÀÌ´Ù. ÀÌ·¸°Ô ºÎ¿©µÈ ±â´ÉÀº over- or under expression µÈ ÇüÁú Àüȯü¸¦ ÅëÇØ È®À뵃 °ÍÀÌ´Ù. (2) Microarray¸¦ ÅëÇØ ½Ä¹°Ã¼ Áý´Ü (plant population)À» screening Çؼ ?olymorphic expression fingerprintÕ¸?ã¾Æ³¾ ¼ö Àִµ¥ ÀÌ·¯ÇÑ Á¤º¸´Â ½Ä¹°Ã¼ÀÇ À¯ÀüÁ¤º¸ (natural accession, transgenic line, mutant, near-isogenic line, substitution line)¿Í ÇÕÇØÁ®¼ ½Ä¹°Ã¼ÀÇ Æ¯Á¤ process¿¡ °ü¿©ÇÏ´Â À¯ÀüÀÚ³ª À¯ÀüÀÚ±ºÀ» ãÀ» ¼ö ÀÖÀ» °ÍÀÌ´Ù (Kehoe et al., 1999). (3) ƯÁ¤¹°ÁúÀÇ Ã³¸® È¿°ú, ¿©·¯ °¡Áö ȯ°æ¿äÀÎ È¿°ú, developmental stage¸¦ microarray¸¦ ÅëÇØ ºÐ¼®ÇÏ¸é ±× °á°ú°¡ quantativeÇÏ°í ¹Ì¼¼ÇÑ À¯ÀüÀÚ º¯È°¡ °¨ÁöµÈ´Ù. ´õ±º´Ù³ª º¯È°¡ ¾ø°Å³ª reduceµÇ´Â À¯ÀüÀÚ±ºµµ µ¿½Ã¿¡ °¨ÁöÇÒ ¼ö ÀÖ´Ù. ÀÌ·¸°Ô ±¤¹üÀ§ÇÑ À¯ÀüÀÚ ¹ßÇö ¾ç»óÀÇ º¯È´Â ½Ä¹°Ã¼ÀÇ ±âº»ÀûÀÎ »ý¸®Çö»óÀÇ °úÁ¤¿¡ ´ëÇÑ ÀÌÇØÀÇ ÆøÀ» ³ÐÈ÷¸ç, heterosisó·³ intractable, complex trait¿¡ ´ëÇÑ ¿¬±¸¸¦ °¡¼ÓȽÃų °ÍÀÌ´Ù (Kehoe et al., 1999). (4) ½Ä¹°Ã¼ÀÇ ¾î¶² Çö»óÀ» ÀÌÇØÇϱâ À§ÇÑ marker À¯ÀüÀÚ·Î »ç¿ëµÇ´Â ¹üÀ§°¡ ÇöÀç´Â »ó´çÈ÷ Á¦ÇѵǾî ÀÖ´Ù. ±×·¯³ª microarray¸¦ ÀÌ¿ëÇÏ¸é »ó´çÈ÷ ¸¹Àº ¼öÀÇ marker À¯ÀüÀÚ¸¦ ¹ß±¼ÇÒ ¼ö ÀÖ´Ù (Schena, 1996). ÀÌ·± marker À¯ÀüÀÚ´Â µ¹¿¬º¯ÀÌü ¹ß±¼À̳ª plant response¸¦ monitorÇϴµ¥ °·ÂÇÑ tool·Î ÀÌ¿ëµÉ ¼ö ÀÖ´Ù. Conclusion ÇöÀç genome sequencing programÀ» ÅëÇØ °è¼Ó ¹ß°ßµÇ´Â sequence informationÀº Á÷Á¢ microarray¿¡ spottingµÇ´Â raw material·Î ±× Á߿伺ÀÌ ÀÖÀ¸¸ç, ¶ÇÇÑ sequence ±× ÀÚü´Â analysis toolÀ» ÀÌ¿ëÇؼ ±â´ÉÀ» ¿¹ÃøÇϴµ¥ ÀÌ¿ëµÈ´Ù. ÀÌ¿¡ ¹ÝÇØ ÃÖ±Ù¿¡ °³¹ßµÈ Microarry´Â Â÷º°È µÈ À¯ÀüÀÚ ¹ßÇöÀ» ºÐ¼®Çؼ ±×µéÀÇ ±â´ÉÀ» ÃßÁ¤ÇÏ´Â tool·Î¼ ÇöÀç °³¹ßµÈ ¹æ¹ý Áß °¡Àå °·ÂÇÑ toolÀÌ´Ù. ÇöÀç plant biologist´Â ¾ÆÁÖ Á¦ÇÑµÈ ºÐ¾ßÀÇ ¿¬±¸¸¦ ÅëÇØ Á¦ÇÑµÈ Á¤º¸¸¦ ¾Ë°í ÀÖ´Ù. ´õ±º´Ù³ª ½Ä¹°Ã¼ÀÇ ¹ß´Þ°ú ¼ºÀåÀº ¾ÆÁÖ º¹ÀâÇÏ°í ÅëÇÕµÈ °úÁ¤ÀÌ´Ù. µû¶ó¼ sequence information°ú microarray informationÀ» ÅëÇÕÇÏ¸é ½Ä¹°Ã¼ ³»¿ÜÀÇ º¯È¿¡ µû¸¥ »ý¸®Àû º¯È¿¡ ´ëÇÑ ÅëÇÕÀûÀÎ ÀÌÇØÀÇ Æ²À» Á¦°øÇØÁØ´Ù. Áö±Ý±îÁö °³¹ßµÈ microarray ºÐ¼®¹ýÀº mRNA transcripts ¼öÁØ¿¡ Á¦ÇѵǾî ÀÖ´Ù. ±×·¯³ª ¿©±â¿¡´Ù ÇöÀç ±Þ¼ÓÈ÷ ¹ßÀüÁßÀÎ proteomics (large scale parallel analysis)³ª protein chipÀÇ Á¤º¸¸¦ ÇÕÇÏ¸é º¸´Ù dzºÎÇÑ »ý¸®ÇÐÀû Á¤º¸¸¦ ¾òÀ» ¼ö ÀÖ´Ù. ¿¹¸¦ µé¸é proteomics´Â microarray¿¡¼ º¼ ¼ö ¾ø´Â ´Ü¹éÁú (gene product)ÀÇ modification±îÁö °¨ÁöÇÒ ¼ö ÀÖ´Ù. µû¶ó¼ Sequence, microarray, proteomics, metabolic profiling µîÀ¸·ÎºÎÅÍ ¾ò¾îÁø ÅëÇÕÀûÀÎ Á¤º¸´Â plant biologists¿¡°Ô ¿©Å±îÁö °æÇèÇÏÁö ¸øÇÑ »õ·Î¿î ¿¬±¸ Á¢±Ù¹ýÀ» ¿ä±¸ÇÏ°í ÀÖ´Ù. References 1. Alon, U., Barkai, N., Notterman, D. 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