EMSA
瀹為獙鎶�鏈強鏂规硶绠�浠?
瀹為獙绠�浠?/p>
鍑濊兌杩佺Щ鎴栫數娉宠縼绉荤巼瀹為獙
锛?/p>
EMSA-electrophoretic mobility shift assay
锛?/p>
鏄竴绉嶇爺绌?/p>
DNA
缁撳悎铔嬬櫧鍜屽叾鐩稿叧鐨?/p>
DNA
缁撳悎搴忓垪鐩镐簰浣滅敤鐨勬妧鏈紝鍙敤浜庡畾鎬у拰瀹氶噺鍒嗘瀽銆傝繖涓�鎶�鏈渶
鍒濈敤浜庣爺绌?/p>
DNA
缁撳悎铔嬬櫧锛?/p>
鐩墠宸茬敤浜庣爺绌?/p>
RNA
缁撳悎铔嬬櫧鍜岀壒瀹氱殑
RNA
搴忓垪鐨勭浉浜掍綔鐢ㄣ�?/p>
閫氬父灏嗙函鍖栫殑铔嬬櫧鍜岀粏鑳炵矖鎻愭恫鍜?/p>
32P
鍚屼綅绱犳爣璁扮殑
DNA
鎴?/p>
RNA
鎺㈤拡涓�鍚屼繚娓╋紝鍦ㄩ潪鍙?/p>
鎬х殑鑱氫笝鐑嚌鑳剁數娉充笂锛屽垎绂诲鍚堢墿鍜岄潪缁撳悎鐨勬帰閽堛�?/p>
DNA-
澶嶅悎鐗╂垨
RNA-
澶嶅悎鐗╂瘮闈炵粨
鍚堢殑鎺㈤拡绉诲姩寰楁參銆?/p>
鍚屼綅绱犳爣璁扮殑鎺㈤拡渚濈爺绌剁殑缁撳悎铔嬬櫧鐨勪笉鍚岋紝
鍙槸鍙岄摼鎴栬�呮槸鍗曢摼銆?/p>
褰?/p>
妫�娴?/p>
濡傝浆褰曡皟鎺у洜瀛愪竴绫荤殑
DNA
缁撳悎铔嬬櫧锛屽彲鐢ㄧ函鍖栬泲鐧斤紝閮ㄥ垎绾寲铔嬬櫧锛屾垨鏍哥粏鑳炴娊
鎻愭恫銆傚湪妫�娴?/p>
RNA
缁撳悎铔嬬櫧鏃讹紝渚濇嵁鐩殑
RNA
缁撳悎铔嬬櫧鐨勪綅缃紝鍙敤绾寲鎴栭儴鍒嗙函鍖栫殑
铔嬬櫧锛屼篃鍙敤鏍告垨鑳炶川缁嗚優鎶芥彁娑层�傜珵浜夊疄楠屼腑閲囩敤鍚泲鐧界粨鍚堝簭鍒楃殑
DNA
鎴?/p>
RNA
鐗囨
鍜屽鏍歌嫹閰哥墖娈碉紙鐗瑰紓锛?/p>
锛?/p>
鍜屽叾瀹冮潪鐩稿叧鐨勭墖娈碉紙闈炵壒寮傦級
锛屾潵纭畾
DNA
鎴?/p>
RNA
缁撳悎铔嬬櫧
鐨勭壒寮傛�с�?/p>
鍦ㄧ珵浜夌殑鐗瑰紓鍜岄潪鐗瑰紓鐗囨鐨勫瓨鍦ㄤ笅锛?/p>
渚濇嵁澶嶅悎鐗╃殑鐗圭偣鍜屽己搴︽潵纭畾鐗瑰紓缁撳悎銆?/p>
鍑濊兌杩佺Щ鎴栫數娉宠縼绉荤巼妫�娴?/p>
锛?/p>
Electrophoretic Mobility Shift Assay, EMSA
锛?/p>
鏄竴绉嶆娴嬭泲
鐧借川鍜?/p>
DNA
搴忓垪鐩镐簰缁撳悎鐨勬妧鏈紝
鏈�鍒濈敤浜庣爺绌?/p>
DNA
缁撳悎铔嬬櫧鍜屽叾鐩稿叧鐨?/p>
DNA
缁撳悎搴忓垪
鐩镐簰浣滅敤锛?/p>
鍙敤浜庡畾鎬у拰瀹氶噺鍒嗘瀽銆?/p>
杩欎竴鎶�鏈?/p>
鐩墠宸茬敤浜庣爺绌?/p>
RNA
缁撳悎铔嬬櫧鍜岀壒瀹氱殑
RNA
搴忓垪鐨勭浉浜掍綔鐢ㄣ�?/p>
鐢变簬寰堝鐮旂┒鐨?/p>
TF
涓嶅叿鏈夌粨鍚堢殑鐗瑰紓鎬э紝鎵�浠ュ嵆浣垮彂鐜?/p>
TF
鍜岃鐮旂┒鐨勫鏍歌嫹閰告帰閽?/p>
缁撳悎锛屼篃涓嶈〃绀哄湪浣撳唴璇?/p>
TF
涓嶈兘鍜屽叾瀹冨鏍歌嫹閰哥粨鍚堬紝杩愮敤涓�浜涚敓鐗╀俊鎭
杞欢锛屽彲浠?/p>
妯℃嫙琛ㄧず
TF
涓庡熀鍥犲惎鍔ㄥ瓙鍖哄鏍歌嫹閰哥粨鍚堢殑鍏蜂綋鎯呭喌锛?/p>
鍙戠幇
TF
鍙互鍜屽惎鍔ㄥ瓙鍖?/p>
PUTATIVE
缁撳悎銆?/p>
鍚屾椂锛岀敱浜?/p>
EMSA
鍦ㄤ綋澶栦笉鑳芥ā
鎷熺粏鑳炲唴浼楀鐢熺墿鍒嗗瓙鐨勭浉浜掍綔鐢紝姣斿锛屾煇涓�
TF
鍦ㄧ粏鑳炲唴鐢变簬鍙楀埌鍏跺畠
TF
鐨勫崗鍚屾垨鑰呬慨楗板悗鎵嶈兘涓庡鏍歌嫹閰哥粨鍚堢殑璇濓紝
閭ｄ箞鍦ㄤ綋澶栨槸涓嶈兘
閲嶇幇杩欎竴缁撴灉鐨勩�?/p>
2003
骞翠笂涓�绡囧叧浜?/p>
ChIP
鏂规硶鐨勪粙缁嶆枃鐚敾鍑?/p>
EMSA
娌℃湁鑰冭檻缁嗚優鍐呭畬鏁磋嚜鐒剁殑鏌撹壊
璐ㄧ粨鏋勫拰璋冭妭鐨勫奖鍝嶈�屽彈鍒板緢澶х殑闄愬埗锛?/p>
涓嬮潰鏄湪涓�涓?/p>
PROTOCAL
1.
鎺㈤拡鐨勬爣璁帮細
(1)
濡備笅璁剧疆鎺㈤拡鏍囪鐨勫弽搴斾綋绯伙細
寰呮爣璁版帰閽?/p>
(1.75pmol/
寰崌
) 2
寰崌
T4 Polynucleotide Kinase Buffer (10X) 1
寰崌
Nuclease-Free Water 5
寰崌
[纬
-32P]A
TP(3,000Ci/mmol at 10mCi/ml) 1
寰崌
T4 Polynucleotide Kinase (5-10u/
寰崌
) 1
寰崌