Ë«·Ö×ÓÓ«¹â»¥²¹¼¼Êõ

ϸ°ûÄÚµ°°×ÖÊÖ®¼äµÄÏ໥×÷ÓûòÐγɵ°°×¸´ºÏÎïÔÚϸ°ûµÄÉúÃü»î¶¯¹ý³ÌÖÐÆð×ÅÖØÒªµÄ×÷Óã¬Ä¿Ç°ÓжàÖÖÊÖ¶ÎÄܹ»ÊµÏÖµ°°×ÖÊÏ໥×÷ÓõÄÑо¿£¬ÈçÃâÒß¹²³Áµí¡¢GST pull down¡¢±íÃæµÈÀë×Ó¹²Õñ¡¢½Íĸ˫ÔÓ½»µÈ¡£±¾ÎÄÖ÷Òª¶ÔÒ»ÖÖ½üÄêÀ´·¢Õ¹ÆðÀ´µÄµ°°×ÖÊÏ໥×÷ÓÃÑо¿·½·¨¡ªË«·Ö×ÓÓ«¹â»¥²¹(bimolecular fluorescence complementation£¬BiFC)¼¼Êõ×öÒ»½éÉÜ¡£

Ë«·Ö×ÓÓ«¹â»¥²¹¼¼ÊõÊÇÒ»ÖÖ»ùÓÚµ°°×ÖÊ»¥²¹¼¼Êõ·¢Õ¹ÆðÀ´µÄ£¬Äܹ»ÓÃÓÚÌåÄÚ»òÌåÍâ¼ì²âµ°°×ÖÊÏ໥×÷Óõļ¼Êõ¡£ÆäʵÑéÔ­ÀíÊǽ«Ó«¹âµ°°×ÔÚºÏÊʵÄλµãÇпª£¬Ðγɲ»·¢Ó«¹âµÄÁ½¸ö¶àëÄ£¨VN¼°VC£©£¬Õâ2¸öƬ¶ÎÔÚϸ°ûÄÚ¹²±í´ï»òÌåÍâ»ìºÏʱ£¬²»ÄÜ×Ô·¢µØ×é×°³ÉÍêÕûµÄÓ«¹âµ°°×ÇÒÔÚ¸ÃÓ«¹âµ°°×µÄ¼¤·¢¹â¼¤·¢Ê±²»ÄܲúÉúÓ«¹â¡£Ëæºó·Ö±ð½«Á½¶Î¶àëÄÓëÓÕ¶üµ°°×ºÍ²¶»ñµ°°×Èںϣ¬Èç¹ûÓÕ¶üµ°°×ºÍ²¶»ñµ°°×ÄÜ·¢ÉúÏ໥×÷Óã¬ÄÇôÁ½¶Î²»ÍêÕûµÄÓ«¹â±¨¸æµ°°×Ƭ¶Î¾Í»á±Ë´Ë¿¿½ü£¬ÖØÐÂÐγɾßÓлîÐÔµÄÓ«¹âµ°°×£¬ÔÚ¸ÃÓ«¹âµ°°×µÄ¼¤·¢¹â¼¤·¢Ê±ÄܲúÉúÓ«¹â¡£×îºóͨ¹ýÓ«¹âÏÔ΢¾µ¹Û²ìÊÇ·ñ´æÔÚÓ«¹â£¬¼´¿ÉÅжÏÓÕ¶üµ°°×Óë²¶»ñµ°°×ÊÇ·ñ´æÔÚÏ໥×÷Óá£

BiTC¼¼ÊõÔ­Àí

BiFCÓÅȱµã

Óŵ㣺

  • Áé»î£º¼ÈÄܹ»ÓÃÓÚÌåÄÚÏ໥×÷ÓõÄÑо¿£¬Ò²Äܹ»ÓÃÓÚÌåÍâÏ໥×÷ÓõÄÑо¿£»
  • ¿ìËÙÖ±¹Û£ºÄܹ»ÔÚÏÔ΢¾µÏÂÖ±½Ó¹Û²ìµ½Ï໥×÷Óýá¹û£»
  • ÊÊÓÃÐԹ㣺BiFCϵͳÒѾ­±»³É¹¦Ó¦ÓÃÓÚ¶¯ÎֲÎï£¬Õæ¾úºÍϸ¾úµÈ²»Í¬µÄËÞÖ÷ϸ°ûÖУ»
  • ±³¾°¸É¾»£¬ÁéÃô¶È¸ß£ºÑéÖ¤½á¹ûÖ»Ðè¼ì²âÓ«¹âµÄÓÐÎÞ£¬±³¾°¸É¾»£¬ÁéÃô¶È¸ß£»
  • Äܹ»¼ì²âÈõÏ໥×÷Óü°Ë²Ê±Ï໥×÷Óã»
  • ¶ÔÒÇÆ÷ÒªÇóµÍ£¬³É±¾¿É¿Ø£¬Êý¾Ý´¦ÀíÏà¶Ô¼òµ¥£»

ȱµã

  • ¶ÔζÈÃô¸Ð£ºÎ¶ȸßʱƬ¶Î¼ä²»Ò×»¥²¹ÐγÉÍêÕûµÄÓ«¹âµ°°×£¬Õâ»á¶ÔÑо¿Ï¸°ûÔÚÉúÀíÌõ¼þϵÄÏ໥×÷ÓôøÀ´¸ºÃæÓ°Ï죨Ŀǰ£¬Ö»ÓлùÓÚvenus¡¢citrineºÍceruleanµÄBiFCϵͳ¿ÉÒÔÔÚÉúÀíζÈÌõ¼þÏÂʵÏÖÆ¬¶Î»¥²¹£©£»
  • ÐÅÏ¢Öͺó£¬ÎÞ·¨ËæÊ±¹Û²ì£ºBiFCϵͳÏëÒª¼ì²âµ½Ó«¹â£¬ÐèÒª¾­¹ýÁ½¸öÓ«¹âµ°°×Ƭ¶Î»¥²¹ÖØÐÂÐγÉÍêÕûµÄ»îÐÔµ°°×£¬ÒÔ¼°Ó«¹âµ°°××ÔÌå´ß»¯Á½¸ö¹ý³Ì£¬¸Ã¹ý³ÌÍùÍùÐèÒª¼¸·ÖÖÓµ½¼¸Ð¡Ê±£¬Òò´Ë¹Û²ìÓ«¹âÐźÅÖͺóÓÚµ°°×ÖʵÄÏ໥×÷Óùý³Ì£¬²»ÄÜʵʱµØ¹Û²ìÏ໥×÷Óùý³Ì£»

¼¼Êõ±³¾°

µ°°×ÖÊ»¥²¹¼¼Êõ

BiFCÊÇ»ùÓÚµ°°×ÖÊ»¥²¹¼¼Êõ·¢Õ¹ÆðÀ´µÄ¡£Ëùνµ°°×ÖÊÆ¬¶Î»¥²¹¼¼Êõ(protein fragment complementation)Êǽ«Ä³¸ö¹¦Äܵ°°×ÇгÉ2¶Î£¬·Ö±ðÓëÁíÍâ2ÖÖÄ¿±êµ°°×ÏàÁ¬ÐγÉ2¸öÈںϵ°°×¡£ÔÚÒ»¸ö·´Ó¦ÌåϵÖУ¬Èç¹û2ÖÖÄ¿±êµ°°×´æÔÚÏ໥×÷Óã¬ÔòʹµÃ2¸ö¹¦Äܵ°°×ÖÊÆ¬¶Î¿¿½ü¡¢»¥²¹²¢Öؽ¨¹¦Äܵ°°×ÖʵĻîÐÔ£¬Í¨¹ý¼ì²â¹¦Äܵ°°×µÄ»îÐÔ¼´Äܹ»ÅжÏÁ½ÖÖÄ¿±êµ°°×ÊÇ·ñ´æÔÚÏ໥×÷Óá£

BiFCÔÚÑØÏ®µ°°×ÖÊÆ¬¶Î»¥²¹¼¼ÊõµÄ»ù´¡ÉϽøÐÐÁËÓÅ»¯£¬µ°°×ÖÊ»¥²¹¼¼ÊõÖй¦Äܵ°°×µÄ»îÐÔÓɵ×Îï·´Ó¦ËùÌåÏÖ£¬Í¨¹ý¼ì²âµ×Îï±ä»¯£¬À´Åжϵ°°×ÖʵÄÏ໥×÷Óã»¶øBiFC¼¼ÊõÔòÊÇÀûÓÃÓ«¹âµ°°×±¾ÉíÄܹ»×ÔÎÒ´ß»¯ÐγÉÓ«¹â»îÐÔÖÐÐIJ¢²úÉúÓ«¹âµ°°×µÄÌØÕ÷¹âÆ×µÄÌØµã£¬Ö±½Ó·´Ó³µ°°×ÖÊÖ®¼äµÄÏ໥×÷Ó㬼¼Êõ¹ý³Ì¸ü¼Ó¼òµ¥£¬½á¹û¸ü¼ÓÖ±¹Û¡£

ÓÃÓÚBiFCϵͳµÄÓ«¹âµ°°×

Ŀǰ³£ÓÃÓÚBiFCϵͳµÄÓ«¹âµ°°×ÓÐGFP(greenfluorescentprotein£¬ÂÌɫӫ¹âµ°°×)¡¢YFP(yellowfluorescentprotein£¬»ÆÉ«Ó«¹âµ°°×)¡¢CFP(cyanfluorescentprotein£¬Çàɫӫ¹âµ°°×)¡¢BFP(bluefluorescentprotein£¬À¶É«Ó«¹âµ°°×)¡¢RFP(redfluorescentprotein£¬ºìɫӫ¹âµ°°×)¡¢Venus¡¢citrine¡¢cerulean¡¢mCherryµÈ¡£

VenusÊÇĿǰ×î³£ÓÃÓÚBiFC·ÖÎöµÄÓ«¹âµ°°×£¬ÒòΪÆäÓ«¹âÇ¿ÇÒ±³¾°Ãô¸Ð¶ÈµÍ£¬ÊÇBiFCϵͳÀíÏëµÄÓ«¹âµ°°×¡£

BiFCϵͳµÄÀ©Õ¹

¶àɫӫ¹â»¥²¹¼¼Êõ

2003Ä꣬Heinemann UµÈÈËÔÚBiFCµÄ»ù´¡ÉϽ¨Á¢ÆðÁ˶àɫӫ¹âµ°°×»¥²¹¼¼Êõ£¨multicolor fluorescent complementation analysis£©¡£ÀûÓöàɫӫ¹âµ°°×»¥²¹¼¼ÊõÄÜͬʱ¼ì²âÒ»¸öϸ°ûÖжà×éµ°°×µÄÏ໥×÷Óã¬ÆäʵÑéÔ­ÀíÊǽ«¶à¸öÓ«¹âµ°°×£¨GFP¡¢CFP¡¢BFPµÈ£©·Ö±ðÔÚÌØ¶¨Î»ÖÃÇпªºóÓëÄ¿±êµ°°×Á¬ÔÚÒ»ÆðÐγÉÈںϵ°°×£¬È»ºó½«ÕâЩÈںϵ°°×»ìºÏ¡£²»Í¬µÄÄ¿±êµ°°×¼ä»á·¢ÉúÏ໥×÷Óã¬Í¬Ê±Ê¹²»Í¬µÄ»¥²¹Ó«¹âµ°°×Ƭ¶Î½áºÏ£¬Ðγɶà¸öµ°°×¸´ºÏÌ壬ͨ¹ýÓ«¹âÏÔ΢¾µ¿É¹Û²ìµ½²»Í¬ÑÕÉ«µÄÓ«¹â£¬´Ó¶øÅж϶à×éµ°°×¼äµÄÏ໥×÷Óá£Í¬Ê±£¬¶àɫӫ¹â»¥²¹¼¼Êõ»¹Äܹ»Í¨¹ýÓ«¹âµÄÇ¿¶È¶Ô²»Í¬×éµ°°×Ö®¼äµÄÏ໥×÷ÓÃÇ¿Èõ½øÐбȽÏ¡£

BiFC-FRET

2008 Ä꣬Shyu YJµÈÈ˽«BiFC¼¼ÊõºÍ FRET ¼¼Êõ½áºÏ£¬½¨Á¢ÁË»ùÓÚË«·Ö×ÓÓ«¹â»¥²¹µÄÓ«¹â¹²ÕñÄÜÁ¿×ªÒƼ¼Êõ(BiFC-FRET)£¬ÀûÓøÃϵͳÄܹ»¼ì²â3¸öµ°°×Ö®¼äµÄÏ໥×÷Óá£BiFC-FRETϵͳÓÉ»ùÓÚvenusµÄBiFCϵͳºÍÇàÉ«µÄÓ«¹âµ°°×cerulean¡£ÆäʵÑéÔ­ÀíΪ£º½«Á½¸öÄ¿±êµ°°×·Ö±ðºÍvenusÓ«¹âµ°°×µÄN£¬C¶ËƬ¶ÎÏàÁ¬£¬Á½Ä¿±êµ°°×µÄÏ໥×÷ÓÃʹµÃvenusµ°°×ÖØ½¨£¨BiFCϵͳ£©¡£½«µÚÈý¸öÄ¿±êµ°°×Óëcerulean¹¹½¨Èںϵ°°×£¬Èç¹û¸ÃÈںϵ°°×Äܹ»ÓëÁ½¸öÄ¿±êµ°°×µÄÒìÔ´¶þ¾ÛÌå·¢ÉúÏ໥×÷Óã¬Ôòcerulean¿ÉÒÔ×÷Ϊ¹©Ì彫ÄÜÁ¿¹²×ªÒÆÖÁÖØ½¨ºóµÄvenusÓ«¹âµ°°×£¬ÊµÏÖ3¸öµ°°×ÖÊÏ໥×÷ÓõĹ²¼ì²â£¨FRET¼¼Êõ£©¡£

BiFC-FRETϵͳ

BiFC-YTH

2011Ä꣬Zheng HYµÈÈ˽«»ùÓÚGFPºÍYFPµÄBiFCϵͳºÍ½Íĸ˫ÔÓ½»£¨YTH£©¼¼ÊõÁªÓÃÀ´Ñо¿Âܲ·»¨Ò¶²¡¶¾ÔÚÄð¾Æ½Íĸ¡¢Ö²ÎïºÍÀ¥³æÏ¸°ûÖеÄ×ÔÎÒÏ໥×÷Óá£BiFC¿ÉÒÔÃÖ²¹YTH¼¼Êõ²»×ãµÄ·½Ã棬ÀýÈçÓÐЩÍâÔ´µ°°×ÔÚ½ÍĸÌåÄÚ´¦ÓÚ²»Í¬µÄλÖ㬼¸ºõûÓпÉÄÜ·¢ÉúÏ໥×÷Ó㬶øBiFC¼¼Êõ¿ÉÖ±½Ó¹Û²âµ½Ï¸°ûÖе°°×ÖÊÎÊÏ໥×÷ÓõÄλÖá£

BiFCϵͳӦÓÃ

Ñо¿µ°°×ÖÊÏ໥×÷ÓÃ

Ñо¿µ°°×ÖÊÏ໥×÷ÓÃÊÇBiFCϵͳ×îÖ÷ÒªÓ¦Óã¬Æù½ñ¸÷ÖÖBiFCϵͳÒѾ­±»³É¹¦ÓÃÓÚ¶àÖÖµ°°×ÖʵÄÏ໥×÷Óã¬ÀýÈçÌåÍâ¡¢²¡¶¾¡¢´ó³¦¸Ë¾ú¡¢½Íĸϸ°û¡¢Ë¿×´Õæ¾ú¡¢²¸È鶯Îïϸ°û¡¢Ö²Îïϸ°û¡¢ÉõÖÁ¸öÌåˮƽµÄµ°°×ÖÊÖ®¼äÏ໥×÷ÓÃÑо¿¡£³ýÁËÄܹ»ÑéÖ¤µ°°×ÖÊÖ®¼äÊÇ·ñ´æÔÚÏ໥×÷Óã¬BiFCϵͳ»¹Äܹ»¶Ôϸ°ûÄÚµ°°×Ï໥×÷ÓÃλÖýøÐÐÑо¿¡£

Ò©Î↑·¢

BiFCÒ²¿ÉÓÃÓÚÒ©Î↑·¢£¬Ê×ÏÈѡȡ¼²²¡Ïà¹ØµÄÖØÒªµ°°×£¬»ùÓÚBiFC¼¼Êõ¿ª·¢BiFCÎÈתϸ°ûÖ꣬²»Í¬µÄÒ©Îï´¦Àíºó¼ì²âÓ«¹âÐźžͿÉɸѡDZÔÚµÄÖÎÁÆÒ©Î¾ßÓÐÁ¼ºÃµÄÓ¦ÓÃǰ¾°¡£

Ïà¹ØÔĶÁ

Á¢¼´Ñ¯¼Û
  • ÐÕÃû

  • µ¥Î»

  • µç»°

  • ×ÉѯÏîÄ¿

  • ÑéÖ¤Âë

  • ÁôÑÔ:

¡¾ÍøÕ¾µØÍ¼¡¿¡¾sitemap¡¿
¡¾ÍøÕ¾µØÍ¼¡¿¡¾sitemap¡¿