鍙嬫儏鎻愮ず锛氭湰绔欐彁渚涘叏鍦�400澶氭墍楂樼瓑闄㈡牎鎷涙敹纰╁+銆佸崥澹爺绌剁敓鍏ュ(xu茅)鑰冭│姝峰勾鑰冪爺鐪熼銆佽€冨崥鐪熼銆佺瓟妗堬紝閮ㄥ垎瀛�(xu茅)鏍℃洿鏂拌嚦2012骞�锛�2013骞�锛涘潎鎻愪緵鏀惰不(f猫i)涓嬭級銆� 涓嬭級娴佺▼锛� 鑰冪爺鐪熼 榛�(di菐n)鎿娾€�鑰冪爺瑭﹀嵎鈥濃€濅笅杓�; 鑰冨崥鐪熼 榛�(di菐n)鎿娾€�鑰冨崥瑭﹀嵎搴�鈥� 涓嬭級
1 婀栧崡甯寖澶у(xu茅)纰╁+鐮旂┒鐢熷叆瀛�(xu茅)鑰冭│鑷懡椤岃€冭│澶х侗 鑰冭│绉戠洰浠g⒓锛歔xxx] 鑰冭│绉戠洰鍚嶇ū锛氭鐗╁垎瀛愮敓鐗╁(xu茅) 锛堟鐗╁(xu茅)灏堟キ(y猫)鍚岀瓑瀛�(xu茅)姝峰姞瑭︾鐩簩锛� 涓€銆佽│鍗风祼(ji茅)妲�(g貌u) 1) 瑭﹀嵎鎴愮妇鍙婅€冭│鏅�(sh铆)闁� 鏈│鍗锋豢鍒嗙偤 100 鍒�锛岃€冭│鏅�(sh铆)闁撶偤 120 鍒嗛悩銆� 2)绛旈鏂瑰紡锛氶枆鍗�銆佺瓎瑭� 3)椤屽瀷绲�(ji茅)妲�(g貌u) a: 鍚嶇ū瑙i噵椤�锛�5 灏忛锛屾瘡灏忛 4 鍒�锛屽叡 20 鍒� b: 鍠爡(xi脿ng)閬告搰椤�锛�5 灏忛锛屾瘡灏忛 2 鍒嗭紝鍏� 10 鍒� c: 濉┖椤�锛屾瘡绌� 1 鍒嗭紝鍏� 20 鍒� d: 绨$瓟椤�锛�5 灏忛锛屾瘡灏忛 6 鍒�锛屽叡 30 鍒� e: 鍟忕瓟椤�, 1-2 灏忛锛屽叡 20 鍒� 浜�銆佽€冭│鍏�(n猫i)瀹硅垏鑰冭│瑕佹眰 1. 妞嶇墿鍩哄洜绲� 鑰冭│鍏�(n猫i)瀹癸細 妞嶇墿鏍稿熀鍥犵祫澶у皬锛岄噸寰�(f霉)搴忓垪锛屽熀鍥犲鏃�锛岀湡鏍稿熀鍥犵殑鏂疯绲�(ji茅)妲�(g貌u)锛涜憠缍犻珨鍩哄洜绲� 鐨勭祼(ji茅)妲�(g貌u)锛岃憠缍犻珨鍩哄洜鐨勭祫鎴愶紝钁夌稜楂斿熀鍥犵殑涓€浜涚祼(ji茅)妲�(g貌u)鐗瑰緛锛涚窔绮掗珨鍩哄洜绲勭殑绲勭箶 绲�(ji茅)妲�(g貌u)锛岀窔绮掗珨鍩哄洜鐨勪竴浜涚壒寰�锛孯NA 绶ㄨ集锛屾鐗╃窗(x矛)鑳炶唱(zh矛)闆勬€т笉鑲茶垏绶氱矑楂旂殑闂�(gu膩n) 绯�锛涢伜鍌充俊鎭湪鍩哄洜绲勪箣闁撶殑娴佸嫊(d貌ng)銆� 鑰冭│瑕佹眰锛� 鎺屾彙鍩哄洜绲�锛屽熀鍥犲鏃忕瓑鍩烘湰姒傚康锛岀悊瑙f鐗╅珨鍏�(n猫i) 3 濂楀熀鍥犵祫鐨勭祼(ji茅)妲�(g貌u)鐗归粸(di菐n)鍙婂叾鐩� 浜掗棞(gu膩n)绯�銆� 2. 妞嶇墿鍙Щ鍕�(d貌ng)鐨勯伜鍌冲洜瀛� 鑰冭│鍏�(n猫i)瀹癸細 2 鍙Щ浣嶅洜瀛愮殑椤炲瀷锛岃綁(zhu菐n)搴у瓙锛孡TR 閫嗚綁(zhu菐n)閷勮綁(zhu菐n)搴у瓙锛岀劇 LTR-鎿綁(zhu菐n)閷勮綁(zhu菐n)搴у瓙锛岃姳妞� 鑺辫憠鐥呮瘨涓殑閫嗚綁(zhu菐n)閷勯叾銆� 鑰冭│瑕佹眰锛氭帉鎻¤綁(zhu菐n)搴у瓙锛屾摤杞�(zhu菐n)搴у瓙鐨勬蹇�锛屾帉鎻″彲绉讳綅鍥犲瓙鐨勯鍨嬶紝浜嗚В鍙Щ浣� 鍥犲瓙鍦ㄥ垎瀛愮敓鐗╁(xu茅)鐮旂┒涓殑鎳�(y墨ng)鐢�銆� 3.鏍搁吀鐨勫垎瀛愭搷浣滃強鍩哄洜閲嶇祫 鑰冭│鍏�(n猫i)瀹癸細 闄愬埗鎬у収(n猫i)鍒囬叾锛屾搷浣滃拰妯�(bi膩o)瑷樻牳閰哥殑閰�锛屽垎瀛愰洔浜�锛岃級楂�锛屽婧� DNA 鑸囪級楂旈噸绲�锛� 闄芥€ч噸绲勯珨鐨勭閬歌垏閼掑畾銆� 鑰冭│瑕佹眰锛� 鎺屾彙闄愬埗鎬у収(n猫i)鍒囬叾锛屽垎瀛愰洔浜わ紝杓夐珨锛孌NA 閲嶇祫绛夊熀鏈蹇�, 鐞嗚В DNA 閲嶇祫鎶€ 琛�(sh霉)鐨勫熀鏈亷绋�銆� 4. PCR 鑰冭│鍏�(n猫i)瀹癸細 PCR 鐨勫熀鏈師鐞�锛孭CR 鐨勫劒(y艒u)鍖�锛岀敱鍩烘湰 PCR 鎷撳睍鐨勭浉闂�(gu膩n)鎶€琛�(sh霉)鍙婂叾鎳�(y墨ng)鐢�銆� 鑰冭│瑕佹眰锛� 鐞嗚В PCR 鐨勫熀鏈師鐞�锛屽強鍏惰鐢熺殑鍏朵粬 PCR,濡� RT-PCR, Tail-PCR, RACE 绛夊父鐢� 鐨� PCR 鏂规硶鍙婃噳(y墨ng)鐢�銆� 5.DNA 鍒嗗瓙妯�(bi膩o)瑷樻妧琛�(sh霉) 鑰冭│鍏�(n猫i)瀹癸細 鍩轰簬 Southern 闆滀氦鐨勫垎瀛愭(bi膩o)瑷樻妧琛�(sh霉)锛岄毃姗�(j墨)寮曠墿鍒嗗瓙妯�(bi膩o)瑷樻妧琛�(sh霉)锛屽簭鍒楁(bi膩o)蹇椾綅榛�(di菐n)鎶€ 琛�(sh霉)锛屽垎瀛愭(bi膩o)瑷樼殑鎳�(y墨ng)鐢ㄩ牁(l菒ng)鍩�銆� 鑰冭│瑕佹眰锛� 鎺屾彙 Southern 闆滀氦, Western 闆滀氦绛夊熀鏈師鐞�锛屾柟娉曞強鎳�(y墨ng)鐢�銆� 6. 鍩哄洜鏂囧韩鍜岀洰鐨勫熀鍥犵殑鍒嗛洟 鑰冭│鍏�(n猫i)瀹癸細 3 鍩哄洜鏂囧韩鐨勬(g貌u)寤猴紝鍏嬮殕宸茬煡鍩哄洜鎴栨牳鑻烽吀搴忓垪淇℃伅鐨勫熀鍥�锛岃泲鐧借唱(zh矛)鍔熻兘浜掕(b菙)鍏嬮殕锛� 瀹氫綅鍏嬮殕锛岄€氶亷 DNA 鍏ㄥ簭鍒楀垎鏋愬厠闅�銆� 鑰冭│瑕佹眰锛� 鎺屾彙鏂囧韩鐨勬(g貌u)寤烘柟娉曪紝浜嗚В甯哥敤鐨勬枃搴鍨�銆� 7. 鍩哄洜鐨勮〃閬�(d谩)瑾�(di脿o)鎺� 鑰冭│鍏�(n猫i)瀹癸細 杞�(zhu菐n)閷勫墠鐨勫熀鍥犳椿鍖栵細鏌撹壊璩�(zh矛)绲�(ji茅)妲�(g貌u)鑸囧熀鍥犳椿鍖栵紝鏍稿熀璩�(zh矛)闄勮憲鍗€(q奴)鑸囧熀鍥犺〃閬�(d谩)锛孌NA 鐢插熀鍖栬(di脿o)鎺у熀鍥犺〃閬�(d谩)鐨勪綔鐢紱杞�(zhu菐n)閷勬按骞崇殑瑾�(di脿o)鎺э細鍩烘湰杞�(zhu菐n)閷勯亷绋�锛岄爢寮忎綔鐢ㄥ厓浠�锛� 鍙嶅紡浣滅敤鍥犲瓙锛岀湡鏍哥敓鐗╁熀鍥犺綁(zhu菐n)閷勮(di脿o)鎺х殑寰�(f霉)闆滄€�锛岀爺绌跺熀鍥犺〃閬�(d谩)瑾�(di脿o)鎺т腑 DNA-铔� 鐧界浉浜掍綔鐢ㄧ殑鏂规硶锛涘熀鍥犺〃閬�(d谩)鐨勮綁(zhu菐n)閷勫悗瑾�(di脿o)鎺э細鐪熸牳鐢熺墿鍓嶉珨 mRNA 鍓垏锛岄珮绛� 妞嶇墿鍏�(n猫i)鍚瓙鐨勭祼(ji茅)妲�(g貌u)锛岄珮绛夋鐗╁壀鍒囬珨铔嬬櫧璩�(zh矛)鐨勭祫鎴愮壒瀹�锛岃畩浣嶅壀鍒�锛孯NA 绶ㄨ集銆� 鑰冭│瑕佹眰锛� 鑰冭│瑕佹眰锛� 鎺屾彙鍩哄洜鍦ㄤ笉鍚屾按骞冲熀鍥犺〃閬�(d谩)瑾�(di脿o)鎺х殑鐗归粸(di菐n)锛屾帉鎻¤畩浣嶅壀鍒�锛孯NA 绶ㄨ集锛岄爢寮忎綔 鐢ㄥ厓浠�锛屽弽寮忎綔鐢ㄥ洜瀛愮瓑鍩烘湰姒傚康锛岀悊瑙h〃瑙€閬哄偝灏嶅熀鍥犺〃閬�(d谩)瑾�(di脿o)鎺х殑褰遍熆銆� 8. 妞嶇墿婵€绱犺垏鍩哄洜琛ㄩ仈(d谩)瑾�(di脿o)鎺� 鑰冭│鍏�(n猫i)瀹癸細 婵€绱犲彈楂旓紝婵€绱犱綔鐢ㄧ殑淇¤櫉鍌抽仦閫斿緫锛屾縺绱犺垏婵€绱犻熆鎳�(y墨ng)鍩哄洜鐨勮〃閬�(d谩)瑾�(di脿o)鎺�锛屼笉鍚屾縺 绱犲皪鍚屼竴鍩哄洜琛ㄩ仈(d谩)鐨勮(di脿o)鎺�銆� 鑰冭│瑕佹眰锛� 鎺屾彙妞嶇墿甯歌鐨勬縺绱犵ó椤炲強婵€绱犲彈楂旂殑鐗归粸(di菐n)銆備簡瑙f鐗╃敘(ch菐n)鐢熸縺绱犵殑閬庣▼鍙婂叾瑾�(di脿o) 鎺э紝浠ュ強婵€绱犺(di脿o)鎺ф鐗╃敓闀风櫦(f膩)鑲茬殑姗�(j墨)鐞�锛岀悊瑙f鐗╂縺绱犱箣闁撶殑鍗�(xi茅)鍚岃垏鎷姉浣滅敤銆� 9. 妞嶇墿鍏夊舰鎱�(t脿i)寤烘垚 鑰冭│鍏�(n猫i)瀹癸細 妞嶇墿鍏夊彈楂旓紝鍏変俊铏熷偝閬為€斿緫锛屽厜瑾�(di脿o)绡€(ji茅)鍩哄洜琛ㄩ仈(d谩)鐨勮綁(zhu菐n)閷勬按骞宠(di脿o)鎺�锛屽厜鏁忚壊绱犲熀鍥� 鐨勮綁(zhu菐n)閷勮(di脿o)鎺х瓑銆� 4 鑰冭│瑕佹眰锛� 浜嗚В妞嶇墿灏嶅厜淇¤櫉鐨勯熆鎳�(y墨ng)閫斿緫銆傛帉鎻″厜鍙楅珨鍙婄稉(j墨ng)鍏哥殑鍏変俊铏熷偝閬為€斿緫銆傜悊瑙e厜瑾�(di脿o) 绡€(ji茅)鍩哄洜琛ㄩ仈(d谩)鐨勮綁(zhu菐n)閷勬按骞宠(di脿o)鎺�锛屽厜鏁忚壊绱犲熀鍥犵殑杞�(zhu菐n)閷勮(di脿o)鎺х瓑銆� 10.楂樼瓑妞嶇墿鐨勯枊鑺辫(di脿o)鎺� 鑰冭│鍏�(n猫i)瀹癸細 闁嬭姳閬庣▼鐨勫熀鍥犺(di脿o)鎺�锛岀挵(hu谩n)澧冨洜瀛愬皪闁嬭姳鏅�(sh铆)闁撴焙瀹氬熀鍥犵殑瑾�(di脿o)鎺с€� 鑰冭│瑕佹眰锛� 鎺屾彙鏄ヨ姳浣滅敤锛屽厜鍛ㄦ湡锛岄枊鑺辩礌绛夊熀鏈蹇�锛屾帉鎻℃鐗╃櫦(f膩)鑲查亷绋嬩腑闁嬭姳璧峰锛岄枊 鑺遍亷绋嬬殑鍩哄洜瑾�(di脿o)鎺э紝浠ュ強鍏夋韩灏嶉枊鑺辨檪(sh铆)闁撴焙瀹氬熀鍥犵殑瑾�(di脿o)鎺х殑褰遍熆銆� 涓�銆佸弮鑰冩浉鐩� [1] 鏇瑰剙妞嶄富绶�. 妞嶇墿鍒嗗瓙鐢熺墿瀛�(xu茅). 楂樼瓑鏁欒偛鍑虹増绀�, 2002 [2] 鏇瑰剙妞嶄富绶�锛屾摤鍗楄姤. 楂樼瓑鏁欒偛鍑虹増绀�, 2004
鍏嶈铂(z茅)鑱叉槑锛氭湰鏂囩郴杞�(zhu菐n)杓夎嚜缍�(w菐ng)绲�(lu貌)锛屽鏈変镜鐘紝璜嬭伅(li谩n)绯绘垜鍊戠珛鍗冲埅闄�锛屽彟锛氭湰鏂囧儏浠h〃浣滆€呭€�(g猫)浜鸿榛�(di菐n)锛岃垏鏈恫(w菐ng)绔欑劇闂�(gu膩n)銆傚叾鍘熷壍(chu脿ng)鎬т互鍙婃枃涓櫝杩版枃瀛楀拰鍏�(n猫i)瀹规湭缍�(j墨ng)鏈珯璀夊(sh铆)锛屽皪鏈枃浠ュ強鍏朵腑鍏ㄩ儴鎴栬€呴儴鍒嗗収(n猫i)瀹�銆佹枃瀛楃殑鐪熷(sh铆)鎬с€佸畬鏁存€�銆佸強鏅�(sh铆)鎬ф湰绔欎笉浣滀换浣曚繚璀夋垨鎵胯锛岃珛璁€鑰呭儏浣滃弮鑰�锛屽苟璜嬭嚜琛屾牳瀵�(sh铆)鐩搁棞(gu膩n)鍏�(n猫i)瀹�銆�
|