¿ÞÂÊ ¸Þ´º ŸÀÌƲ À̹ÌÁö
¿¬±¸µ¿Çâ

Home < ¿­¸°¸¶´ç < ¿¬±¸µ¿Çâ

       ÇÕ¼º »ý¹°Çп¡¼­ÀÇ ¹«¼¼Æ÷ ´Ü¹éÁú ÇÕ¼º ½Ã½ºÅÛÀÇ ÀÀ¿ë(Ãæ³²´ë ±èµ¿¸í ±³¼ö, ÀÌ°æÈ£ ¹Ú»ç)
       dmkim@cnu.ac.kr        
       ±èµ¿¸í        2013.05.06 17:12        20125
´Ù¿î·Îµå : ÇÕ¼º »ý¹°Çп¡¼­ÀÇ ¹«¼¼Æ÷ ´Ü¹éÁú ÇÕ¼º ½Ã½ºÅÛÀÇ ÀÀ¿ë(Ãæ³²´ë ±èµ¿¸í ±³¼ö, ÀÌ°æÈ£ ¹Ú»ç).pdf(592 Kb)
 

ÇÕ¼º »ý¹°Çп¡¼­ÀÇ ¹«¼¼Æ÷ ´Ü¹éÁú ÇÕ¼º ½Ã½ºÅÛÀÇ ÀÀ¿ë

 

 

 

             ÀÌ°æÈ£ ¹Ú»ç, ±èµ¿¸í ±³¼ö

                                                                                                        Ãæ³²´ëÇб³

 

1. °³¿ä

 

ÇÕ¼º »ý¹°ÇÐÀº »õ·Î¿î »ý¹°ÇÐÀû ½Ã½ºÅÛÀÇ ±¸ÇöÀ» À§ÇÑ »ý¹°ÇÐÀû ºÎÇ°µéÀÇ ÇÕ¼º, ¿£Áö´Ï¾î¸µ ±×¸®°í Á¶ÇÕ¿¡ ±âÃʸ¦ µÎ°í ÀÖ´Ù. DNA, RNA±×¸®°í ´Ü¹éÁú µîÀÇ »ý¹°ÇÐÀû ºÎÇ°µé·Î »õ·Ó°Ô ¼³°èµÈ ¸ðµâµéÀº ´Ü°èÀû È®ÀåÀ» ÅëÇØ ¸ñÀû »ê¹°ÀÇ ÃÖÀûÈ­µÈ »ý»êÀ» À§ÇÑ ½Ã½ºÅÛÀ¸·Î À̾îÁø´Ù. ¼¼Æ÷ ³»¿¡¼­ ´ëºÎºÐÀÇ »ý¹°ÇÐÀû ±â´É°ú È­ÇÐ ¹ÝÀÀ µîÀ» ¼öÇàÇÏ´Â ´Ü¹éÁúÀº ÇÕ¼º»ý¹°ÇÐ ½Ã½ºÅÛÀ» ±¸¼ºÇÏ´Â ±âº»ÀûÀÎ ±¸¼º ¿ä¼Ò ÁßÀÇ ÇϳªÀÌ´Ù. ÇÑÆí, »ý¹°ÇÐÀû ºÎÇ°À¸·Î »ç¿ëµÉ ¼ö ÀÖ´Â ´Ü¹éÁúµéÀ» ¾ÏȣȭÇÏ´Â À¯ÀüÀÚ ¼­¿­Àº À¯ÀüÀÚ ¼­¿­ºÐ¼®±â¼úÀÇ ±Þ¼ÓÇÑ ¹ßÀü°ú ÇÔ²² ±âÇϱ޼öÀûÀ¸·Î Áõ°¡ÇÏ°í ÀÖÀ¸³ª ÀÌ¿¡ ºñÇÏ¿© ´Ü¹éÁúÀÇ ±¸Á¶ ºÐ¼® ¹× ±â´ÉÀ» ¹àÈ÷´Â ÀÛ¾÷Àº À¯ÀüÁ¤º¸ÀÇ Áõ°¡¼Óµµ¸¦ µû¸£Áö ¸øÇÏ°í ÀÖ´Ù (Figure 1). ÀÌ´Â ±âÁ¸ÀÇ ¼¼Æ÷ ±â¹ÝÀÇ ´Ü¹éÁú ÇÕ¼º¹ýÀ¸·Î À¯ÀüÁ¤º¸¸¦ ¹ø¿ªÇÏ´Â ¼Óµµ ¹× ¾²·çDzÀÇ ÇÑ°è¿¡ ±âÀÎÇÑ´Ù°í ÇÒ ¼ö ÀÖÀ¸¸ç, »ý¹°ÇÐÀû ºÎÇ°ÀÇ °í¼Ó°³¹ßÀ» À§Çؼ­´Â ´ë¾ÈÀûÀÎ ´Ü¹éÁú ÇÕ¼º¹ýÀÌ ÇÊ¿äÇÑ ½ÇÁ¤ÀÌ´Ù.1ÀÌ·¯ÇÑ ¸Æ¶ô¿¡¼­ ¹«¼¼Æ÷ ´Ü¹éÁú ÇÕ¼º ½Ã½ºÅÛÀº »ì¾Æ ÀÖ´Â ¼¼Æ÷ÀÇ »ç¿ë¿¡¼­ ºñ·ÔµÇ´Â ¼¼Æ÷ ±â¹Ý ´Ü¹éÁú ÇÕ¼º¹ýÀÇ ÇѰ踦 ¹þ¾î³ª, À¯ÀüÀÚ ¹ßÇöÀÇ ºÐÀÚ»ý¹°ÇÐÀû Á¢±ÙÀ» ¿ëÀÌÇÏ°Ô Çϵµ·Ï ´Ù¾çÇÑ ÇüÅÂÀÇ À¯¿¬¼ºÀ» Á¦°øÇÑ´Ù. º» ¶õ¿¡¼­´Â À¯Àü Á¤º¸ÀÇ ¹ø¿ª¿¡¸¸ ±¹ÇѵÇÁö ¾Ê°í ¸ñÀû ´Ü¹éÁúÀÇ 3Â÷ ±¸Á¶ Çü¼º ¹× À¯Áö¸¦ À§ÇÏ¿© ´Ù¾çÇÏ°Ô Ç¥ÁØÈ­µÈ ¹«¼¼Æ÷ ´Ü¹éÁú ÇÕ¼º¹ýÀ» ¼Ò°³ÇÏ°í, ´õ ³ª¾Æ°¡ ¹«¼¼Æ÷ ´Ü¹éÁú ÇÕ¼º ½Ã½ºÅÛÀÌ ´Ù¸¥ ¿µ¿ªÀÇ °úÇÐ ºÐ¾ß¿¡¼­ ÇàÇØÁö´Â ´Ù¾çÇÑ ½ÇÇè ±â¹ýµé°úÀÇ Á¶ÇÕÀ» ÅëÇÏ¿© °Ô³ð ¼­¿­ÀÇ ½Å¼ÓÇÑ ±â´É ºÐ¼®À» À§ÇÑ »õ·Î¿î °³³äÀÇ ´Ü¹éÁú ÇÕ¼º ¹× ºÐ¼® ½Ã½ºÅÛÀ¸·Î ¹ßÀüµÇ´Â ³»¿ëÀ» ´ã°í ÀÖ´Â ¿¬±¸µéÀ» ¼Ò°³ÇÏ°í °íÂûÇÏ°íÀÚ ÇÑ´Ù.

 

Figure 1. Widening gap between the amounts of sequence information (red circles)

                           and of protein structures (blue circles).

 

2. ¸ñÀû ´Ü¹éÁúÀÇ Æ¯¼º¿¡ µû¶ó Ç¥ÁØÈ­µÈ ¹«¼¼Æ÷ ´Ü¹éÁú ÇÕ¼º ½Ã½ºÅÛ

 

¹«¼¼Æ÷ ´Ü¹éÁú ÇÕ¼º ½Ã½ºÅÛÀº »ì¾Æ ÀÖ´Â ¼¼Æ÷¸¦ ÀÌ¿ëÇÑ ¼¼Æ÷ ±â¹ÝÀÇ ÇÕ¼º¹ý°ú Â÷º°È­µÈ Á¢±Ù ¹æ½ÄÀ» äÅÃÇÏ°í ÀÖ´Ù. ±âº»ÀûÀ¸·Î ¼¼Æ÷ ±â¹ÝÀÇ ´Ü¹éÁú ÇÕ¼º¹ýÀº ¸ñÀû ´Ü¹éÁúÀ» ¾ÏȣȭÇÏ´Â À¯ÀüÀÚ°¡ ÇüÁúÀüȯµÈ ¼¼Æ÷¸¦ ¹è¾çÇÏ°í ¼¼Æ÷ Æļ⠰úÁ¤À» °ÅÃÄ ¼ö°ÅÇÏ¿© ´Ü¹éÁúÀ» È®º¸ÇÑ´Ù. ¹Ý¸é¿¡ ¹«¼¼Æ÷ ´Ü¹éÁú ÇÕ¼º¹ýÀº ¹è¾çµÈ ¼¼Æ÷¸¦ ÆļâÇÏ¿© ¸®º¸¿ÀÁ»À» Æ÷ÇÔÇÑ ´Ü¹éÁú ÇÕ¼º ±â±¸µéÀÌ Á¸ÀçÇÏ´Â ¼¼Æ÷ ÃßÃâ¾×À» ¿ì¼±ÀûÀ¸·Î È®º¸ÇÏ°í, ÁغñµÈ ¼¼Æ÷ ÃßÃâ¾×¿¡ À¯ÀüÀÚ¿Í ÇÔ²² Ãß°¡ÀûÀ¸·Î ¾Æ¹Ì³ë»ê, ¿¡³ÊÁö¿ø ¹× ¿ÏÃæ¿ë¾× µîÀ» ÇÔ²² ÷°¡ÇÏ¿© ¸ñÀû ´Ü¹éÁúÀ» ÇÕ¼ºÇϵµ·Ï ÇÑ´Ù. »ì¾Æ ÀÖ´Â ¼¼Æ÷¸¦ »ç¿ëÇÏÁö ¾Ê°í, ¼¼Æ÷·ÎºÎÅÍ ÃßÃâµÈ ´Ü¹éÁú »ýÇÕ¼º ±â±¸¸¦ ÀÌ¿ëÇÏ¿© ¼¼Æ÷ ¹Ù±ù¿¡¼­ ´Ü¹éÁúÀ» »ý»êÇÏ´Â ¹«¼¼Æ÷ ´Ü¹éÁú ÇÕ¼º ½Ã½ºÅÛÀÇ °¡Àå Å« Ư¡Àº ÀÎÀ§ÀûÀ¸·Î ÇÕ¼º Á¶°ÇÀ» Á¶ÀÛÇÒ ¼ö ÀÖ´Ù´Â °ÍÀÌ´Ù. ¼¼Æ÷ÀÇ »ýÀå¿¡ ÇÊ¿äÇÑ º¹ÀâÇÑ ´ë»ç ³×Æ®¿öÅ©ÀÇ À¯Áö°¡ ÇÊ¿äÇÏÁö ¾Ê±â ¶§¹®¿¡ ´Ü¹éÁú ÇÕ¼º¿¡ ¿µÇâÀ» ¹ÌÄ¡Áö ¾Ê´Â ´Ù¸é ¼¼Æ÷ µ¶¼ºÀ» °®°Å³ª Ä¡¸íÀûÀÎ ¿µÇâÀ» ÁÖ´Â È­ÇÕ¹°ÀÇ »ç¿ëÀÌ °¡´ÉÇÏ´Ù.

 

ÀÎÀ§ÀûÀ¸·Î ¹ÝÀÀ Á¶°ÇÀ» Á¶ÀÛÇÒ ¼ö ÀÖ´Â ¹«¼¼Æ÷ ´Ü¹éÁú ÇÕ¼º ½Ã½ºÅÛÀÇ ´ëÇ¥ÀûÀÎ ¸ðµ¨·Î´Â Ueda¿¬±¸ÆÀ¿¡ ÀÇÇØ °³¹ßµÈ ¡°PURE (Protein synthesis Using Recombinant Elements)¡± ½Ã½ºÅÛÀ» µé ¼ö ÀÖ´Ù.2 ÀÌ ¿¬±¸ÆÀÀº ´ëÀå±ÕÀÇ 31°¡ÁöÀÇ ´Ü¹éÁú »ýÇÕ¼º ¹ø¿ª ÀÎÀÚµéÀ» °³º°ÀûÀ¸·Î ÁغñÇÏ°í, 46Á¾ tRNA, ¸®º¸Á¶¿È ¹× ´Ü¹éÁú ÇÕ¼º ±âÁúµéÀ» µû·Î ÁغñÇÑ ÈÄ ¸ñÀû À¯ÀüÀÚ¿ÍÀÇ È¥ÇÕ¹°À» ¸¸µé¾î ´Ü¹éÁúÀ» ÇÕ¼ºÇÒ ¼ö ÀÖ´Â ½Ã½ºÅÛÀ» ±¸ÃàÇÏ¿´´Ù. ÀÌ·¯ÇÑ PURE ½Ã½ºÅÛÀº °¢°¢ÀÇ ÀÎÀÚµéÀ» ½±°Ô ´ëüÇÒ ¼ö Àֱ⠶§¹®¿¡ ¸ñÀû ´Ü¹éÁú ÇÕ¼º»Ó¸¸ÀÌ ¾Æ´Ï¶ómRNA display, ºñõ¿¬ ¾Æ¹Ì³ë»êÀÇ µµÀÔ µî¿¡ ÀûÇÕÇÏ°Ô °³¼±µÇ¾î È°¿ëµÇ±âµµ ÇÏ¿´´Ù. ÇÏÁö¸¸, ´Ü¹éÁú ¹ßÇö ÀÎÀÚµéÀÇ °³º°ÀûÀÎ ¹ßÇö ¹× ºÐ¸®/Á¤Á¦ °úÁ¤ÀÌ ÇʼöÀûÀÎ PURE ½Ã½ºÅÛÀº ½ºÄÉÀÏ-¾÷À» ÅëÇÑ ¸ñÀû ´Ü¹éÁúÀÇ ´ë·® »ý»ê¿¡´Â ÀûÇÕÇÏÁö ¾Ê´Ù.

 

³·Àº ¼öÀ²ÀÇ ÇѰ踦 °®´Â PURE½Ã½ºÅÛ°ú ºñ±³ÇÏ¿© ¼¼Æ÷ ÃßÃâ¾×À» »ç¿ëÇÏ´Â ¹«¼¼Æ÷ ´Ü¹éÁú ÇÕ¼º ½Ã½ºÅÛÀº ¼ö mg/mlÀÇ ¹ßÇö·®À¸·Î ³ôÀº ¼öÀ²·Î ´Ü¹éÁúÀ» ÇÕ¼ºÇÒ ¼ö ÀÖ´Â ÀåÁ¡À» °¡Áö°í ÀÖÀ¸¸ç,3 ÀÌ·¯ÇÑ ½Ã½ºÅÛÀ» ÀÌ¿ëÇÏ¿© ¸ñÀû ´Ü¹éÁúÀÇ ³ôÀº ¹ßÇö ¼öÀ²À» À¯ÁöÇϸ鼭µµ ¸ñÀû¿¡ ¸Â°Ô ´Ü¹éÁú ÇÕ¼º ÀÎÀÚµéÀ» À籸¼ºÇÏ°íÀÚ ÇÏ´Â ¿¬±¸µéÀÌ ÁøÇàµÇ¾î¿Ô´Ù. ¼¼Æ÷ ÃßÃâ¾×¿¡ Á¸ÀçÇÏ´Â tRNA¸¦ Á¦°ÅÇÏ¿© À¯ÀüÀÚÀû ¸®ÇÁ·Î±×·¡¹ÖÀ» ÀÌ¿ëÇÏ¿© ´Ü¹éÁúÀ» ¿£Áö´Ï¾î¸µ Çϰųª,4-6 ¼¼Æ÷ Æļâ¾×³»ÀÇ ÀúÇØ ÀÎÀÚ¸¦ ¼±ÅÃÀûÀ¸·Î Á¦°ÅÇÏ¿© ´Ü¹éÁú ÇÕ¼º È¿À²À» Áõ°¡½ÃÅ°°íÀÚ ÇÏ´Â ½ÃµµµéÀÌ ÁøÇàµÇ¾î¿ÔÀ¸¸ç,7 ÀÌ·¸µí ¸ñÀû¿¡ ¸Â°Ô ´Ü¹éÁú ÇÕ¼º ¹ÝÀÀ ȯ°æÀ» Á¶ÀýÇÒ ¼ö ÀÖ´Â ¹«¼¼Æ÷ ´Ü¹éÁú ÇÕ¼º ½Ã½ºÅÛÀº ´õ ³ª¾Æ°¡ ¸ñÀû ´Ü¹éÁúÀÇ 3Â÷ ±¸Á¶¸¦ °í·ÁÇÏ¿© ¾ÈÁ¤ÀûÀÎ Á¢ÈûÀ» À¯µµÇÏ°íÀÚ ´Ù¾çÇÑ ÇüÅ·ΠȰ¼ºÇü ´Ü¹éÁú ÇÕ¼ºÀ» À§ÇÑ Ç¥ÁØÈ­°¡ ÁøÇàµÇ¾ú´Ù.

 

´Ü¹éÁú ¹ßÇö ¼öÀ²À» ³ôÀÌ´Â ½Ãµµ¿Í ´õºÒ¾î È¿°úÀûÀÎ ´Ü¹éÁúÀÇ Á¢Èû°ú ¹ø¿ª ÈÄ ¼ö½Ä°úÁ¤À» À§ÇÏ¿© Ç¥ÁØÈ­µÈ ¹«¼¼Æ÷ ´Ü¹éÁú ÇÕ¼º ½Ã½ºÅÛÀÌ °³¹ßµÇ¾ú´Ù (Figure 2). ´Ü¹éÁúÀÇ Á¢ÈûÀº ±× ±â´É°ú ¹ÐÁ¢ÇÑ °ü°è¸¦ °¡Áö°í ÀÖ°í, ȯ¿ø Á¶°ÇÀÇ ´ëÀå±Õ À¯·¡ÀÇ ¹«¼¼Æ÷ ´Ü¹éÁú ÇÕ¼º ½Ã½ºÅÛÀº ÀÌȲȭ°áÇÕÀ» °®´Â ¸ñÀû ´Ü¹éÁúÀÇ ÇÕ¼ºÀ» Á¦ÇÑÇÑ´Ù. ÀÌȲȭ°áÇÕÀº ÁÖ·Î ¼¼Æ÷Áú ±¸È¹¿¡¼­ ÀϾ´Âµ¥, ¿øÇÙ¼¼Æ÷¿¡¼­´Â periplasmÀ̳ª ÁøÇÙ¼¼Æ÷ÀÇ endoplasmic reticulumÀÇ °ü»ó¿¡¼­ ÀϾ´Ù. ÇÏÁö¸¸, ¹«¼¼Æ÷ ´Ü¹éÁú ÇÕ¼º¹ý¿¡¼­´Â º»ÁúÀûÀ¸·Î ÀÌȲȭ°áÇÕ Çü¼ºÀ» ¹æÇØÇÏ´Â ¿ä¼ÒµéÀÌ Á¸ÀçÇϸç, À̸¦ ÇØ°áÇϱâ À§ÇÏ¿© ¼¼Æ÷ ÃßÃâ¾× ³»ÀÇ ÀÌȲȭ°áÇÕÀÇ È¯¿øÀ» À¯µµÇÏ´Â ´Ü¹éÁúµéÀ» ºñÈ°¼ºÈ­½ÃÅ°°í, »êÈ­/ȯ¿ø Æ÷ÅÙ¼ÈÀ» Á¶ÀýÇÏ¿© ÀÌȲȭ°áÇÕÀ» ¸ñÀû ´Ü¹éÁú¿¡ µµÀÔÇÒ ¼ö ÀÖÀ¸¸ç, ±× °á°ú ´Ü¹éÁúÀÇ È°¼º ¿ª½Ã Áõ°¡ÇÏ´Â °ÍÀ» È®ÀÎÇÒ ¼ö ÀÖ¾ú´Ù.8-11

 

¹«¼¼Æ÷ ´Ü¹éÁú ÇÕ¼º ½Ã½ºÅÛ¿¡¼­ ¸·´Ü¹éÁúÀÇ °¡¿ëÀûÀÎ ¹ßÇöÀ» À§ÇÏ¿© ´Ü¹éÁúÀÇ ±¸Á¶Àû ¾ÈÁ¤¼ºÀ» ³ôÀÌ°íÀÚ °è¸éÈ°¼ºÁ¦ ȤÀº ¸®Æ÷¿ÀÁ»°ú °°Àº ÀÎÀ§ÀûÀÎ ±â±¸µéÀ» »ç¿ëÇϱ⵵ ÇÑ´Ù.12-14 Áï, ¸·´Ü¹éÁúÀÇ °ú¹ßÇöÀº ¼¼Æ÷ÀÇ »ýÀåÀ» ÀúÇØÇÏ´Â ¿ªÇÒÀ» Çϱ⠽±Áö¸¸, ¹«¼¼Æ÷ ´Ü¹éÁú ÇÕ¼ºÀº ±Ù¿øÀûÀ¸·Î ÀÌ·¯ÇÑ ¹®Á¦¸¦ ÇÇÇÒ ¼ö ÀÖ´Ù. ÇÏÁö¸¸, ¸·´Ü¹éÁúÀÇ ³ôÀº ¼Ò¼ö¼º¿¡ ÀÇÇÏ¿© ÀÀÁýü·Î ¹ßÇöµÇ´Â °æ¿ì°¡ ´ëºÎºÐÀ̸ç, ÀÌ·¯ÇÑ ¹®Á¦¸¦ ÇØ°áÇÏ°íÀÚ °è¸éÈ°¼ºÁ¦ ¶Ç´Â Áö¹æ»êÀ¸·Î ÀÌ·ç¾îÁø ¸®Æ÷¿ÀÁ»ÀÇ Ã·°¡´Â ¼¼Æ÷ ³»¿¡¼­ÀÇ ¸·´í¹éÁúÀÇ ±¸Á¶¸¦ À¯ÁöÇÒ ¼ö ÀÖ´Â ÁöÁöü·Î ÀÛ¿ëÇÏ¿© ¸·´Ü¹éÁúÀÇ °¡¿ë¼ºÀ» ³ôÀ̸鼭 È°¼ºÇü ¸·´Ü¹éÁúÀ» ÇÕ¼ºÇÒ ¼ö ÀÖ´Â ½Ã½ºÅÛÀÌ °³¹ßµÇ¾ú´Ù.

 

¸¶Áö¸·À¸·Î ¹«¼¼Æ÷ ´Ü¹éÁú ÇÕ¼ºÀÌ ÁøÇàµÇ´Â µ¿¾È ¹ø¿ª ÈÄ ¼ö½ÄÀÌ °¡´ÉÇÑ ½Ã½ºÅÛÀÌ ±¸ÃàµÇ¾ú´Ù.15 ´ëÀå±ÕÀ̳ª ¹Ð ¹è¾Æ À¯·¡ÀÇ ¼¼Æ÷ ÃßÃâ¾×Àº ¼÷ÁÖ°¡ °®´Â Ư¼ºÀ» ÀÌ¾î ¹Þ±â ¶§¹®¿¡ ´Ü¹éÁú ³»·Î ´çÀÇ ¼ö½ÄÀÌ °¡´ÉÇÏÁö ¾Ê´Ù.16 À̸¦ ÇØ°áÇÏ°íÀÚ RRL (rabbit reticulocyte), insect cell, Hela cell µîÀÇ ´Ù¾çÇÑ ÁøÇÙ¼¼Æ÷¸¦ ÀÌ¿ëÇÏ¿© ¼¼Æ÷ ÃßÃâ¾×À» ¸¶·ÃÇÏ¿© ´ç ´Ü¹éÁú ÇÕ¼ºÀ» °¡´ÉÇÏ°Ô ÇÏ¿´´Ù.17 RRL ±â¹ÝÀÇ ¹«¼¼Æ÷ ´Ü¹éÁú ÇÕ¼º ½Ã½ºÅÛÀº »ó´ëÀûÀ¸·Î ¸Å¿ì ³·Àº ´Ü¹éÁú ÇÕ¼º ¼öÀ²À» °®±â ¶§¹®¿¡ ½Ç¿ë¼ºÀÌ ¶³¾îÁö°í insect cellÀÇ °æ¿ì¿¡´Â ÇÕ¼º ¼öÀ²Àº »ó´ëÀûÀ¸·Î ³ôÁö¸¸ ´ç ¼ö½Ä È¿À²ÀÌ ³·Àº ÆíÀÌ´Ù. ÀÌ¿¡ ¹ÝÇØ human cell lineÀ¸·ÎºÎÅÍ ÁغñµÈ ¼¼Æ÷ ÃßÃâ¾×À» »ç¿ëÇÒ °æ¿ì¿¡´Â RRL, insect ¼¼Æ÷ ±â¹ÝÀÇ ½Ã½ºÅÛ¿¡ ºñÇÏ¿© ´Ü¹éÁú ÇÕ¼º ¼öÀ²µµ ³ôÀ» »Ó¸¸ ¾Æ´Ï¶ó ´ç ¼ö½ÄÀÇ È¿À²µµ °¡Àå ³ô±â ¶§¹®¿¡ ´ç ´Ü¹éÁúÀÇ ÇÕ¼º¿¡ ÀûÇÕÇÑ ½Ã½ºÅÛÀ¸·Î °³¹ßµÇ¾ú´Ù.18

Figure 2. Cell-free protein synthesis coupled with various

   post-translational modification reactions

 

3. ¹Ì¼¼À¯Ã¼Ç÷§Æû ±â¹ÝÀÇ ¹«¼¼Æ÷ ´Ü¹éÁú ÇÕ¼º½Ã½ºÅÛ

 

¹«¼¼Æ÷ ´Ü¹éÁú ÇÕ¼º ½Ã½ºÅÛÀº ¸ñÀû¿¡ ¸Â°Ô ¹ÝÀÀ Á¶¼ºÀ» Á¶ÀÛÇÒ ¼ö ÀÖÀ» »Ó¸¸ ¾Æ´Ï¶ó, ¹ÝÀÀ±â ÇüÅ¿¡ ±¸¾Ö ¹ÞÁö ¾Ê°í ÀÎÀ§ÀûÀÎ ¹ÝÀÀ ȯ°æ¿¡¼­ ´Ü¹éÁú ÇÕ¼ºÀ» ¼öÇàÇÒ ¼ö ÀÖ´Ù. Áï, ¹«¼¼Æ÷ ´Ü¹éÁú ÇÕ¼º ½Ã½ºÅÛÀÇ °¡Àå Å« ÀåÁ¡Àº ¼¼Æ÷ ¹è¾çÀ̳ª Ŭ·Î´×°ú °°Àº ´Ü°è¸¦ ¹èÁ¦Çϸ鼭 À¯Àü Á¤º¸¸¦ ƯÁ¤ °ø°£¿¡ ±¸¾Ö ¹ÞÁö ¾Ê°í ¹ø¿ªÇÒ ¼ö ÀÖ´Â °ÍÀÌ´Ù. ƯÈ÷, ¸¶ÀÌÅ©·ÎÇ÷çÀ̵ñ½º ½Ã½ºÅÛ°úÀÇ Á¶ÇÕÀ¸·Î ´Ü¹éÁú ÇÕ¼º°ú ºÐ¼®À» À§ÇÑ »õ·Î¿î Ç÷§ÆûÀ¸·Î ¹ßÀüµÇ¾î ¿Ô´Ù (Figure 3). ¹Ì¼¼À¯Ã¼ ½Ã½ºÅÛÀ¸·Î ÁغñµÈ ¼ö¹é ³ª³ë¸®ÅÍ ¼öÁØÀÇ °ø°£¿¡¼­ CECFÇü½ÄÀ¸·Î ´Ü¹éÁú ÇÕ¼ºÀ» ÁøÇà ÇÔÀ¸·Î½á ¼ö ¹é °¡ÁöÀÇ À¯ÀüÀÚ¸¦ µ¿½Ã/º´·ÄÀûÀ¸·Î ÃÊ°í¼Ó ¹ßÇöÀ» °¡´ÉÇÏ°Ô ÇÏ´Â ½Ã½ºÅÛÀÌ °³¹ßµÇ¾ú´Ù.19-21 ´õ ³ª¾Æ°¡ Quake ¿¬±¸ÆÀÀº ´Ü¹éÁú-´Ü¹éÁú »óÈ£ÀÛ¿ë ¿¬±¸¿¡ »ç¿ëÇÒ ¼ö ÀÖ´Â ¹«¼¼Æ÷ ´Ü¹éÁú ÇÕ¼º¹ý ±â¹ÝÀÇ ¹Ì¼¼À¯Ã¼ ½Ã½ºÅÛÀ» °³¹ßÇÏ¿´´Ù.22 À¯¸® ½½¶óÀ̵å Å©±âÀÇ ¹Ì¼¼À¯Ã¼ ¹ÝÀÀ±â¿¡¼­ ¾à 14,792°³ÀÇ ¹ÝÀÀÀ» µ¿½Ã¿¡ ÁøÇàÇÒ ¼ö ÀÖ´Â ÁýÀûµµ¸¦ °®Ãß¾úÀ¸¸ç, À¯ÀüÀڷκÎÅÍ ¹ßÇöµÈ ´Ü¹éÁúµéÀÌ ¹Ì¸® ÁغñµÈ Ç×ü¿¡ °íÁ¤ÀÌ µÇ°í 43 Á¾·ùÀÇ Streptococcus pneumonia À¯·¡ÀÇ ´Ü¹éÁúµéÀÌ ÀÛÀº Åë·Î¸¦ ÅëÇÏ¿© ±³Â÷¹ÝÀÀÀÌ ÁøÇàµÈ´Ù. ±× ÈÄ, ´Ü¹éÁú-´Ü¹éÁú »óÈ£ÀÛ¿ëÀÌ ÀϾ ´Ü¹éÁú ½ÖÀ» °°Àº ȸ·Î¿¡¼­ È®ÀÎÇÒ ¼ö ÀÖ¾úÀ¸¸ç, ±âÁ¸ÀÇ ¿¬±¸¿¡¼­ ¾Ë·ÁÁø »óÈ£ÀÛ¿ëÇÏ´Â ´Ü¹éÁú ½Ö ÀÌ¿Ü¿¡µµ »õ·Î¿î Á¾·ùÀÇ ´Ü¹éÁú-´Ü¹éÁú »óÈ£ÀÛ¿ëÀ» ÀÏÀ¸Å°´Â ´Ü¹éÁú ½ÖÀ» ã¾Æ³¾ ¼ö ÀÖ¾ú´Ù. ÀÌ·¸°Ô ¸¶ÀÌÅ©·ÎÇ÷çÀ̵ñ½º ½Ã½ºÅÛÀº ÃÊ°í¼Ó/º´·Ä½Ä ´Ü¹éÁú ÇÕ¼ºÀ» °¡´ÉÇÏ°Ô Çϴ°¡ ÇÏ¸é ´õ ³ª¾Æ°¡ ¶Ç ´Ù¸¥ ºÐ¼® ½Ã½ºÅÛ°úÀÇ Á¶ÇÕÀ» ÅëÇÏ¿© »õ·Î¿î ºÐ¼® Ç÷§ÆûÀ» Á¦°øÇϱ⵵ ÇÏ¿´´Ù. Seefeld ¿¬±¸ÆÀÀº ¸¶ÀÌÅ©·ÎÇ÷çÀ̵ñ½º ½Ã½ºÅÛÀ» surface plasmon resonance (SPR) Ĩ À§¿¡¼­ ±¸ÇöÇÏ¿© ¹«¼¼Æ÷ ´Ü¹éÁú ÇÕ¼ºÀ» ÅëÇÑ ´Ü¹éÁúÀÇ ÇÕ¼º °úÁ¤À» ½Ç½Ã°£À¸·Î ºÐ¼®ÇÒ ¼ö ÀÖ´Â ½Ã½ºÅÛÀ» °³¹ßÇÏ¿´´Ù.23 ÀÌ·¯ÇÑ ºÐ¼® ½Ã½ºÅÛÀº ´Ü¹éÁú-´Ü¹éÁú, ´Ü¹éÁú-DNA ¶Ç´Â ´Ü¹éÁú-RNA µîÀÇ »óÈ£ÀÛ¿ëÀ» ½Ç½Ã°£À¸·Î ºÐ¼®ÇÒ ¼ö ÀÖ´Â »õ·Î¿î ¹«¼¼Æ÷ ´Ü¹éÁú ÇÕ¼º¹ý ±â¹ÝÀÇ ºÐ¼® ½Ã½ºÅÛÀ¸·Î »ç¿ëµÉ ¼ö ÀÖ´Ù. 

 

Figure 3. Cell-free protein synthesis based microfluidics system.

                          A: A cell-free protein synthesis system in a microfluidic device using passive pumping

                          B: An in vitro microfluidic platform for highthroughput screening of protein interactions,      

                              called protein interaction network generator (PING)

                          C: A single-step biosensing platform combined cell-free protein synthesis with SPRi.

 

4. ¹«¼¼Æ÷ ´Ü¹éÁú ÇÕ¼º ½Ã½ºÅÛ ±â¹ÝÀÇ in situ ´Ü¹éÁú ¾î·¹ÀÌ

 

´Ü¹éÁú ¾î·¹ÀÌ ±â¼úÀº ¼ö¹é¿¡¼­ ¼öõ °³ÀÇ ´Ü¹éÁúÀ» µ¿½Ã¿¡ ºÐ¼® °¡´ÉÇÏ°Ô Çϸ鼭 ½Ã°£À» ´ÜÃà ½ÃÅ°°í ºñ¿ëÀ» Àý°¨Çϵµ·Ï ÇÑ´Ù. ÇÏÁö¸¸, ±âÁ¸ÀÇ ¼¼Æ÷ ±â¹ÝÀÇ ´Ü¹éÁú ÇÕ¼º¹ýÀ¸·Î À¯ÀüÀÚ Å¬·Î´×, ¼¼Æ÷ ¹è¾ç, ´Ü¹éÁú ºÐ¸® ¹× Á¤Á¦¸¦ °ÅÃÄ ´Ü¹éÁú ¾î·¹À̸¦ À§ÇÏ¿© ´Ü¹éÁúÀ» ÁغñÇÏ´Â µ¥¿¡´Â ¸¹Àº ½Ã°£°ú ³ëµ¿·ÂÀÌ ¹ß»ýÇÑ´Ù. À̸¦ ÇØ°áÇϱâ À§ÇÑ ´ë¾ÈÀûÀÎ ¹æ¹ýÀ¸·Î ¹«¼¼Æ÷ ´Ü¹éÁú ÇÕ¼º¹ý ±â¹ÝÀÇ in situ ´Ü¹éÁú ¾î·¹ÀÌ ±â¹ýÀÌ °³¹ßµÇ¾î ¿Ô´Ù. Taussig¿¬±¸ÆÀÀº ¸ñÀû ´Ü¹éÁúÀ» ÀνÄÇÏ´Â Ç×ü°¡ ¹Ù´Ú¿¡ °íÁ¤µÈ 24 well Çü½ÄÀÇ plate¿¡¼­ À¯ÀüÀÚ¿Í ÇÔ²² ¹«¼¼Æ÷ ´Ü¹éÁú ÇÕ¼º¿¡ ÇÊ¿äÇÑ ¿ä¼ÒµéÀ» ÇÔ²² ¹ÝÀÀÇÏ¿© ´Ü¹éÁúÀ» °íÁ¤ÇÏ´Â ¡®protein in situ array (PISA)¡¯ °³³äÀ» µµÀÔÇÏ¿´´Ù.24, 25 À¯ÀüÀÚ ¾î·¹ÀÌ¿Í ´Þ¸® ´Ü¹éÁú ¾î·¹ÀÌ´Â º¯¼ºÀÌ ÀϾ±â ½±±â ¶§¹®¿¡ Àå±â º¸Á¸ÀÌ Èûµé´Ù. ÀÌ·¯ÇÑ PISA ±â¹ýÀº ÇÊ¿äÇÒ ¶§¿¡ ¼ö ½Ã°£ ³»¿¡ ´Ü¹éÁú ¾î·¹À̸¦ ÁغñÇÒ ¼ö Àֱ⠶§¹®¿¡ º¸°ü ¹®Á¦¸¦ ÇÇÇÒ ¼ö ÀÖ´Ù. ´Ü¹éÁú ¾î·¹ÀÌ°¡ DNA ¾î·¹ÀÌ ±â¹ý°ú ÇÔ²² »ç¿ëµÇ¸é¼­ well plate Çü½ÄÀÌ ¾Æ´Ñ DNA ¾î·¹À̷κÎÅÍ ´Ü¹éÁú ¾î·¹ÀÌ·Î ÀüȯÇÏ´Â ±â¹ýµéÀÌ À̾ °³¹ßµÇ¾ú´Ù. LaBaer ¿¬±¸ÆÀÀº ¸ÕÀú DNA ¾î·¹À̸¦ ÁغñÇÑ ÈÄ ¹«¼¼Æ÷ ´Ü¹éÁú ÇÕ¼º µ¿¾È ¹ßÇöµÈ ¿¡ÇÇÅéÀ» °®´Â ¸ñÀû ´Ü¹éÁúÀÌ º»·¡ÀÇ DNA ¾î·¹ÀÌ ±Ùó¿¡ °íÁ¤µÇ´Â ½ÄÀÇ ¡®Nucleic acid programmable protein array (NAPPA)¡¯ ±â¼úÀ» °³¹ßÇÏ¿´´Ù.26, 27ÀÌ·¯ÇÑ ±â¼úÀº DNA ¾î·¹À̷κÎÅÍ ¹«¼¼Æ÷ ´Ü¹éÁú ÇÕ¼º¹ýÀ» ÅëÇÏ¿© ´Ü¹éÁú ¾î·¹ÀÌ·Î ÀüȯÇÏ´Â °³³äÀ» µµÀÔÇÑ °ÍÀ¸·Î PISAÀÇ ±â¹ý¿¡ ºñÇÏ¿© µ¿½Ã¿¡ ó¸®ÇÒ ¼ö ÀÖ´Â À¯ÀüÀÚÀÇ ¼ö¸¦ Å©°Ô Çâ»ó ½Ãų ¼ö ÀÖ¾ú´Ù. ±× ÈÄ, À¯¸® ½½¶óÀ̵忡¼­ ¹«¼¼Æ÷ ´Ü¹éÁú ÇÕ¼º¹ýÀ» ±â¹ÝÀ¸·Î ´Ü¹éÁú ¾î·¹ÀÌÀÇ ÁýÀûµµ¸¦ Å©°Ô Çâ»ó½ÃÅ°´Â ±â¹ýÀÌ ¸¶·ÃµÇ¾ú´Ù. Hoheisel ¿¬±¸ÆÀÀº ¡®multiple spotting technique (MST)¡¯À» ÅëÇÏ¿© À¯¸® ½½¶óÀÌµå ´ç 13,000°³ÀÇ ´ÞÇÏ´Â ´Ü¹éÁú ¾î·¹À̸¦ ¸¶·ÃÇÒ ¼ö ÀÖµµ·Ï ÇÏ¿´´Ù.28 Ni-NTA°¡ °íÁ¤µÈ ½½¶óÀ̵忡 ¸ÕÀú À¯ÀüÀÚ ¾î·¹À̸¦ Çü¼º ½ÃÅ°°í À¯ÀüÀÚ°¡ ¾î·¹ÀÌ µÇ¾ú´ø µ¿ÀÏÇÑ À§Ä¡¿¡ ´Ü¹éÁú ÇÕ¼ºÀ» À§ÇÑ È¥ÇÕ¾×À» ´Ù½Ã ¾î·¹ÀÌ ÇÏ¿© ¹«¼¼Æ÷ ´Ü¹éÁú ÇÕ¼º ¹ÝÀÀÀÌ ÁøÇàµÇ°í ÃÖÁ¾ÀûÀ¸·Î ¸ñÀû ´Ü¹éÁúÀÌ ¹ßÇöµÇ¾î °íÁ¤µÇµµ·Ï ÇÏ¿´´Ù.

Figure 6. In situ protein array methods based on a cell-free protein synthesis system. 

 

5. °á·Ð ¹× °íÂû

 

¹«¼¼Æ÷ ´Ü¹éÁú ÇÕ¼º ½Ã½ºÅÛÀº À¯ÀüÀÚ Å¬·Î´×, ¹è¾ç°øÁ¤ µî ¼¼Æ÷ ±â¹ÝÀÇ ´Ü¹éÁú ÇÕ¼º ½Ã½ºÅÛÀÌ °®´Â ÇѰ踦 ±Øº¹ÇÏ°í, ´Ù·®ÀÇ ´Ü¹éÁú ºÎÇ°µéÀ» °í¼ÓÀ¸·Î ÇÕ¼º ÇÒ ¼ö ÀÖ´Â ±â¼ú·Î »ç¿ëµÉ ¼ö ÀÖ´Ù. ƯÈ÷, ¼¼Æ÷¸·ÀÇ Á¸Àç ¾øÀÌ ±ÕÀÏÇÑ ¾×»ó¹ÝÀÀ ÇüÅ·Π´Ü¹éÁúÀ» ÇÕ¼ºÇÒ ¼ö ÀÖ´Ù´Â ¹«¼¼Æ÷ ´Ü¹éÁú ÇÕ¼º±â¼úÀÇ ÀåÁ¡Àº ÀÎÇØ ¹«¼¼Æ÷ ´Ü¹éÁú ¹ßÇö ½Ã½ºÅÛÀº ´Ü¼øÈ÷ À¯ÀüÁ¤º¸¸¦ ´Ü¹éÁú·Î ¹ø¿ªÇÏ´Â µµ±¸·Î¼­ÀÇ ±â´ÉÀ» ¶Ù¾î³Ñ¾î, ÀΰøÀûÀÎ ÇÕ¼º ¹× ºÐ¼® Ç÷§Æû°úÀÇ À¶ÇÕÀ» ÅëÇØ À¯ÀüÀÚ ¼­¿­»ó¿¡ ÀÔ·ÂµÈ ±â´ÉÀ» È¿À²ÀûÀ¸·Î Çؼ®ÇÏ°í Á¶ÀÛÇÒ ¼ö ÀÖ´Â ±â¼úÀû ±â¹ÝÀ» Á¦°øÇÑ´Ù. ÃÖ±Ù µé¾î, º»·Ð¿¡¼­ ¾ð±ÞµÈ »ç·ÊµéÀ» Æ÷ÇÔÇÏ¿© ¼¼Æ÷ÀÇ ¹ÙÀÌ¿ÀÇÕ¼º±â±¸¸¦ À¯ÀüÀÚ ¹ßÇöÀÌ ¿ä±¸µÇ´Â ´Ù¾çÇÑ ÀÀ¿ëºÐ¾ß¿¡ Àû¿ëÇÏ°íÀÚ ÇÏ´Â ¿¬±¸°¡ È°¹ßÈ÷ ÁøÇàµÇ°í ÀÖÀ¸¸ç, ¹«¼¼Æ÷ ´Ü¹éÁú ÇÕ¼º ±â¼úÀÇ È°¿ë¹üÀ§´Â ÇâÈÄ Áö¼ÓÀûÀ¸·Î È®´ëµÉ °ÍÀ¸·Î ¿¹»óµÈ´Ù.

 

Âü°í¹®Çå

 

1. Harris, D.C. and Jewett, M.C. Cell-free biology: exploiting the interface between synthetic biology and synthetic chemistry. Current Opinion in Biotechnology 23, 672-678 (2012).
2. Shimizu, Y., Inoue, A., Tomari, Y., Suzuki, T., Yokogawa, T., Nishikawa, K. and Ueda, T. Cell-free translation reconstituted with purified components. Nature Biotechnology  19, 751-755 (2001).
3. Kim, T.W., Keum, J.W., Oh, I.S., Choi, C.Y., Kim, H.C. and Kim, D.M. An economical and highly productive cell-free protein synthesis system utilizing fructose-1,6-bisphosphate as an energy source. Journal of Biotechnology  130, 389-393 (2007).
4. Jackson, R.J., Napthine, S. and Brierley, I. Development of a tRNA-dependent in vitro translation system. RNA  7, 765-773 (2001).
5. Kanda, T., Takai, K., Yokoyama, S. and Takaku, H. Knocking out a specific tRNA species within unfractionated Escherichia coli tRNA by using antisense (complementary) oligodeoxyribonucleotides. FEBS Letters  440, 273-276 (1998).
6. Kanda, T., Takai, K., Yokoyama, S. and Takaku, H. An easy cell-free protein synthesis system dependent on the addition of crude Escherichia coli tRNA. Journal of Biochemistry  127, 37-41 (2000).
7. Michel-Reydellet, N., Woodrow, K. and Swartz, J. Increasing PCR fragment stability and protein yields in a cell-free system with genetically modified Escherichia coli extracts. Journal of Molecular Microbiology and Biotechnology  9, 26-34 (2005).
8. Goerke, A.R. and Swartz, J.R. Development of cell-free protein synthesis platforms for disulfide bonded proteins. Biotechnology and Bioengineering 99, 351-367 (2008).
9. Kim, D.M. and Swartz, J.R. Efficient production of a bioactive, multiple disulfide-bonded protein using modified extracts of Escherichia coli. Biotechnology and Bioengineering  85, 122-129 (2004).
10. Oh, I.S., Kim, D.M., Kim, T.W., Park, C.G. and Choi, C.Y. Providing an oxidizing environment for the cell-free expression of disulfide-containing proteins by exhausting the reducing activity of Escherichia coli S30 extract. Biotechnology Progress 22, 1225-1228 (2006).
11. Yin, G. and Swartz, J.R. Enhancing multiple disulfide bonded protein folding in a cell-free system. Biotechnology and Bioengineering 86, 188-195 (2004).
12. Bannwarth, M. and Schulz, G.E. The expression of outer membrane proteins for crystallization. Biochimica et Biophysica Acta 1610, 37-45 (2003).
13. Junge, F., Haberstock, S., Roos, C., Stefer, S., Proverbio, D., Dotsch, V. and Bernhard, F. Advances in cell-free protein synthesis for the functional and structural analysis of membrane proteins. New Biotechnology 28, 262-271 (2011).
14. Schwarz, D., Klammt, C., Koglin, A., Lohr, F., Schneider, B., Dotsch, V. and Bernhard, F. Preparative scale cell-free expression systems: new tools for the large scale preparation of integral membrane proteins for functional and structural studies. Methods 41, 355-369 (2007).
15. Endo, Y. and Sawasaki, T. Cell-free expression systems for eukaryotic protein production. Current Opinion in Biotechnology  17, 373-380 (2006).
16. Takai, K., Sawasaki, T. and Endo, Y. The wheat-germ cell-free expression system. Current Pharmaceutical Biotechnology  11, 272-278 (2010).
17. Ezure, T., Suzuki, T., Shikata, M., Ito, M., Ando, E., Utsumi, T., Nishimura, O. and Tsunasawa, S. Development of an insect cell-free system. Current Pharmaceutical Biotechnology  11, 279-284 (2010).
18. Zeenko, V.V., Wang, C., Majumder, M., Komar, A.A., Snider, M.D., Merrick, W.C., Kaufman, R.J. and Hatzoglou, M. An efficient in vitro translation system from mammalian cells lacking the translational inhibition caused by eIF2 phosphorylation. RNA 14, 593-602 (2008).
19. Angenendt, P., Nyarsik, L., Szaflarski, W., Glokler, J., Nierhaus, K.H., Lehrach, H., Cahill, D.J. and Lueking, A. Cell-free protein expression and functional assay in nanowell chip format. Analytical Chemistry  76, 1844-1849 (2004).
20. Kinpara, T., Mizuno, R., Murakami, Y., Kobayashi, M., Yamaura, S., Hasan, Q., Morita, Y., Nakano, H., Yamane, T. and Tamiya, E. A picoliter chamber array for cell-free protein synthesis. Journal of Biochemistry  136, 149-154 (2004).
21. Khnouf, R., Beebe, D.J. and Fan, Z.H. Cell-free protein expression in a microchannel array with passive pumping. Lab Chip  9, 56-61 (2009).
22. Gerber, D., Maerkl, S.J. and Quake, S.R. An in vitro microfluidic approach to generating protein-interaction networks. Nature Methods  6, 71-74 (2009).
23. Seefeld, T.H., Halpern, A.R. and Corn, R.M. On-chip synthesis of protein microarrays from DNA microarrays via coupled in vitro transcription and translation for surface plasmon resonance imaging biosensor applications. J Am Chem Soc 134, 12358-12361 (2012).
24. He, M. and Taussig, M.J. Single step generation of protein arrays from DNA by cell-free expression and in situ immobilisation (PISA method). Nucleic Acids Res 29, e73-73 (2001).
25. He, M. and Taussig, M.J. DiscernArray technology: a cell-free method for the generation of protein arrays from PCR DNA. J Immunol Methods 274, 265-270 (2003).
26. Ramachandran, N., Hainsworth, E., Bhullar, B., Eisenstein, S., Rosen, B., Lau, A.Y., Walter, J.C. and LaBaer, J. Self-assembling protein microarrays. Science 305, 86-90 (2004).
27. Ramachandran, N., Hainsworth, E., Demirkan, G. and LaBaer, J. On-chip protein synthesis for making microarrays. Methods Mol Biol 328, 1-14 (2006).
28. Angenendt, P., Kreutzberger, J., Glokler, J. and Hoheisel, J.D. Generation of high density protein microarrays by cell-free in situ expression of unpurified PCR products. Mol Cell Proteomics 5, 1658-1666 (2006).




Total:118 page:(8/5)
54 Á¤º¸ ÀÌ¿µÈÆ RNA ±¸Á¶ ±â¹Ý ¼¼Æ÷³» ¿Âµµ°¨ÀÀ¼¾¼­ 15.08.21 12081
53 Á¤º¸ ÃÖÁ¾Çö ¹Ì»ý¹°À» ÀÌ¿ëÇÑ ÀÌŸÄÜ»ê »ý»ê ¹× µ¿Çâ 15.06.30 13833
52 Á¤º¸ ÀÌÆòõ Whole-Genome and Whole-Population ºÐ¼®À» ÀÌ¿ë.. 15.05.27 11954
51 Á¤º¸ ³ëÁ¤Çý ¼¼±ÕÀÇ ¹ø¿ª ½ºÆ®·¹½º ¹ÝÀÀ 15.05.27 11035
50 Á¤º¸ À±¿©ÁØ »ýÇÕ¼º°ú À¯±âÇÕ¼º À¶ÇÕÀ» ÅëÇÑ Ãµ¿¬¹° ÀüÇÕ¼º ±â.. 15.04.30 12533
49 Á¤º¸ À¯º´Á¶ ¹ÙÀÌ¿ÀÀ̼ҺÎź¿Ã(Bio-isobutanol)ÀÇ »ý»ê±â¼ú°ú .. 15.04.30 17244
48 Á¤º¸ ±è¼±¿ø ¹Ì»ý¹° ¼¼Æ÷°øÀå ´ë»çÁ¶Àý ±â¼úÀÇ °³¹ßµ¿Çâ 15.03.19 12273
47 Á¤º¸ À̱չΠÁö´ÉÇü ¹ÙÀÌ¿À ½Ã½ºÅÛ¿¡ ÀÇÇÑ °íÇ°Áú Ç×ü ÀǾàÇ°.. 15.03.16 14986
46 Á¤º¸ ÀÌ»ó¿± ÇÕ¼º Á¶Àý sRNAs °³¹ß ¹× ´ë»ç°øÇп¡¼­ÀÇ ÀÀ¿ë¿¡ .. 15.03.16 13116
45 Á¤º¸ Á¶ÁÖÇö »ýü¹æ¾îÆéŸÀ̵åÀÇ ´ë·®»ý»ê ¹× ½Ç¿ëÈ­¿¡ ´ëÇÑ .. 15.02.25 19157
44 Á¤º¸ ±èµ¿¸³ ¹Ì»ý¹° Àü±â»ýÇÕ¼ºÀ» À§ÇÑ ÁöÁöü °³¹ß µ¿Çâ 15.02.25 11506
43 Á¤º¸ ±è±ÙÁß °íºÎ°¡°¡Ä¡ ´Ü¹éÁú ¼ÒÀ縦 ¾ÏȣȭÇÑ ³­¹ßÇö À¯Àü.. 15.01.27 15101
42 Á¤º¸ À̼º±¹ Evolutionary adaptation ¹× reverse engineering.. 15.01.27 13278
41 Á¤º¸ ±èÇÊ ±³ ÀΰøÀûÀÎ »êÈ­½ºÆ®·¹½º³»¼º ÄÚ¸®³×¹ÚÅ׸®¿ò 14.12.10 13358
40 Á¤º¸ ÇÑÁøÈñ Áø¼¼³ë»çÀ̵带 ÀÌ¿ëÇÑ ³úÀÎÁö±â´É Çâ»ó¿¡ ´ëÇÑ .. 14.12.09 15391
39 Á¤º¸ Á¤±âÁØ ÄÚ¸®³×¹ÚÅ׸®¿ò ±â¹Ý ÇÕ¼º»ý¹°ÇÐÀû ¿¬±¸ µ¿Çâ 14.11.21 16440
[1] [2] [3] [4] [5] [6] [7] [8]