{"id":3191,"date":"2018-07-27T14:20:09","date_gmt":"2018-07-27T06:20:09","guid":{"rendered":"http:\/\/si.secda.info\/yb_mdn\/?page_id=3191"},"modified":"2018-07-27T14:20:09","modified_gmt":"2018-07-27T06:20:09","slug":"ylex%e9%85%b5%e6%af%8d%e8%a1%a8%e9%81%94%e8%a9%a6%e5%8a%91%e7%9b%92","status":"publish","type":"page","link":"https:\/\/si.secda.info\/yb_mdn\/?page_id=3191","title":{"rendered":"YLEX\u9175\u6bcd\u8868\u9054\u8a66\u5291\u76d2"},"content":{"rendered":"
\n
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\n
\n
\n
\n

YLEX\u9175\u6bcd\u8868\u9054\u8a66\u5291\u76d2
\u63cf\u8ff0<\/strong>
\u57fa\u65bcINRA INAPG\u8a31\u53ef\u5c08\u5229*\u7684YLEX Expression Kit\u63d0\u4f9b\u4e86\u4e00\u7a2e\u7c21\u55ae\u7684\u65b9\u6cd5\uff0c\u7528\u65bc\u514b\u9686\u548c\u8868\u9054\u9175\u6bcd\uff0c\u89e3\u8102\u8036\u6c0f\u9175\u6bcd\u4e2d\u611f\u8208\u8da3\u7684\u57fa\u56e0\u3002\u4f7f\u7528\u8a72\u8a66\u5291\u76d2\uff0c\u901a\u904e\u9078\u64c7\u63d0\u4f9b\u7684\u8868\u9054\u8f09\u9ad4pYLEX1\u6216pYLSC1\uff0c\u53ef\u4ee5\u5728\u7d30\u80de\u5167\u8868\u9054\u9ad8\u6c34\u5e73\u7684\u7570\u6e90\u86cb\u767d\u8cea\u6216\u5f9e\u7d30\u80de\u5206\u6ccc\u5230\u57f9\u990a\u57fa\u4e2d\u3002<\/p>\n

YLEX\u4e2d\u63d0\u4f9b\u7684\u8f09\u9ad4\u542b\u6709\u5f37\u96dc\u5408\u555f\u52d5\u5b50\uff0c\u5176\u651c\u5e36\u4f86\u81eapXPR2\u7684\u56db\u500b\u4e32\u806f\u62f7\u8c9d\u7684\u4e0a\u6e38\u6fc0\u6d3b\u5b50\u5e8f\u5217\uff08UAS1B\uff09\u7247\u6bb5\u548c\u6700\u5c0f\u7684pLEU2\u7247\u6bb5\u3002\u8207\u5e38\u7528\u7684\u8036\u6c0f\u9175\u6bcd\u5c6c\u555f\u52d5\u5b50\uff08pXPR2\uff09\u4e0d\u540c\uff0c\u9019\u7a2e\u7a69\u5b9a\u7684\u96dc\u5408\u555f\u52d5\u5b50\u4e0d\u65b7\u5730\u6307\u5c0e\u86cb\u767d\u8cea\u8868\u9054\uff0c\u800c\u4e0d\u53d7\u57f9\u990a\u57fa\u4e2d\u71df\u990a\u548c\u74b0\u5883\u56e0\u7d20\u7684\u591a\u91cd\u5f71\u97ff\u3002<\/p>\n

\u7576\u5177\u6709\u96dc\u5408\u555f\u52d5\u5b50\u7684\u96a8\u5f8c\u88ab\u514b\u9686\u7684\u76ee\u7684\u57fa\u56e0\u69cb\u5efa\u7684\u8cea\u7c92\u88ab\u9078\u64c7\u7684\u9650\u5236\u9176\u7dda\u6027\u5316\u6642\uff0c\u5b83\u8b8a\u6210\u53ef\u4ee5\u901a\u904e\u8f49\u5316\u904e\u7a0b\u4e2d\u7684\u540c\u6e90\u91cd\u7d44\u6574\u5408\u5230\u89e3\u8102\u8036\u6c0f\u9175\u6bcd\u57fa\u56e0\u7d44\u4e2d\u7684\u8868\u9054\u76d2\u3002\u53d6\u6c7a\u65bc\u5206\u6ccc\u4fe1\u865f\u662f\u5426\u51fa\u73fe\u5728\u8cea\u7c92\u4e0a\uff0c\u6210\u529f\u7684\u8f49\u5316\u9ad4\u6e96\u5099\u597d\u8868\u9054\u6216\u5206\u6ccc\u91cd\u7d44\u86cb\u767d\u3002\u6709\u95dc\u8a73\u7d30\u4fe1\u606f\uff0c\u8acb\u95b1\u8b80\u672c\u76ee\u9304\u4e2d\u5f15\u7528\u7684\u6587\u7ae0\u3002<\/p>\n

* INRA\uff08\u570b\u5bb6\u8fb2\u696d\u7814\u7a76\u6240\uff09\u548cINAPG\uff08Institut National Agronomique Paris-Grignon\uff09<\/p>\n

\"\"<\/p>\n

\"\"<\/p>\n

\u7279\u5fb5<\/strong>
\u00a0 \u00a0 \u2022 \u5b89\u5168\uff1a
\u00a0 \u00a0 \u00a0 Y. lipolytica\u88ab\u7f8e\u570bFDA\uff08\u98df\u54c1\u548c\u85e5\u54c1\u7ba1\u7406\u5c40\uff09\u6b78\u985e\u70baGRAS\uff08\u901a\u5e38\u8a8d\u70ba\u662f\u5b89\u5168\u7684\uff09
\u00a0 \u00a0 \u2022 \u7c21\u55ae\uff1a
\u00a0 \u00a0 \u00a0\u8868\u9054\u7570\u6e90\u86cb\u767d\u8cea\u7684\u7c21\u55ae\u5de5\u5177
\u00a0 \u00a0 \u2022 \u5bb9\u6613\u64cd\u4f5c\uff1a
\u00a0 \u00a0 \u00a0\u50cf\u5927\u8178\u687f\u83cc\u548c\u91c0\u9152\u9175\u6bcd\u4e00\u6a23
\u00a0 \u00a0 \u2022 \u7a69\u5b9a\uff1a
\u00a0 \u00a0 \u00a0\u8f09\u9ad4\u548c\u69cb\u5efa\u7684\u8cea\u7c92\u5177\u6709\u5f88\u5f37\u7684\u7a69\u5b9a\u6027
\u00a0 \u00a0 \u2022 \u53ef\u9760\uff1a
\u00a0 \u00a0 \u00a0\u6574\u5408\u5728\u57fa\u56e0\u7d44\u4e2d\u76f8\u540c\u4f4d\u9ede\u7684\u8f09\u9ad4
\u00a0 \u00a0 \u2022 \u9748\u6d3b\uff1a
\u00a0 \u00a0 \u00a0\u63d0\u4f9b\u8868\u9054\u548c\u5206\u6ccc\u8f09\u9ad4\uff08\u86cb\u767d\u8cea\u53ef\u4ee5\u5728\u7d30\u80de\u5167\u8868\u9054\u6216\u5f9e\u7d30\u80de\u5206\u6ccc\u5230\u57f9\u990a\u57fa\u4e2d\uff09
\u00a0 \u00a0 \u2022 \u9ad8\u589e\u9577\u80fd\u529b\uff1a
\u00a0 \u00a0 \u00a0\u5206\u6ccc\u80fd\u529b\u9ad8\uff0c\u7522\u91cf\u9ad8
\u00a0 \u00a0 \u2022 \u6e1b\u5c11\u86cb\u767d\u8cea\u964d\u89e3\uff1a
\u00a0 \u00a0 \u00a0\u6c92\u6709\u7531\u7279\u6b8a\u86cb\u767d\u9176\u7f3a\u9677\u578b\u8036\u6c0f\u9175\u6bcd\u83cc\u682a\u5408\u6210\u7684\u7d30\u80de\u5916\u86cb\u767d\u9176
\u00a0 \u00a0 \u2022 \u5927\u91cf\u751f\u7522\uff1a
\u00a0 \u00a0 \u00a0\u91cd\u7d44\u86cb\u767d\u7684\u5de5\u696d\u5927\u898f\u6a21\u751f\u7522
\u00a0 \u00a0 \u2022 \u8f03\u5c11\u7684\u9ad8\u7cd6\u57fa\u5316\uff1a
\u00a0 \u00a0 \u00a0\u8207\u91c0\u9152\u9175\u6bcd\u4e0d\u540c\uff0c\u5b83\u80fd\u5920\u9032\u884c\u8907\u96dc\u86cb\u767d\u8cea\u7684\u7ffb\u8b6f\u5f8c\u52a0\u5de5<\/p>\n

\u6b65\u9a5f<\/p>\n

\"\"<\/p>\n

\u61c9\u7528<\/strong>
\u7d30\u80de\u5167\u6216\u7d30\u80de\u5916\u7684\u7570\u6e90\u86cb\u767d\u8cea\u8868\u9054\u53d6\u6c7a\u65bcGRAS\u9175\u6bcd\u4e2d\u7684\u9078\u5b9a\u8f09\u9ad4\u3002
\u8036\u6c0f\u9175\u6bcd\u8f09\u9ad4
\u8a72\u8a66\u5291\u76d2\u4e2d\u5305\u62ec\u5169\u7a2e\u8f09\u9ad4\uff08pYLEX1\u548cpYLSC1\uff09\uff0c\u5b83\u5011\u53ef\u7528\u65bc\u89e3\u8102\u8036\u6c0f\u9175\u6bcd\u4e2d\u611f\u8208\u8da3\u7684\u86cb\u767d\u8cea\u7684\u7d30\u80de\u5167\u8868\u9054\u6216\u5206\u6ccc\u3002\u4e00\u822c\u800c\u8a00\uff0c\u5982\u679c\u6a19\u9776\u86cb\u767d\u662f\u80de\u8cea\u548c\u975e\u7cd6\u57fa\u5316\u7684\uff0c\u5247pYLEX1\u8f09\u9ad4\u662f\u66f4\u597d\u7684\u9078\u64c7\u3002\u5982\u679c\u60a8\u611f\u8208\u8da3\u7684\u86cb\u767d\u8cea\u901a\u5e38\u662f\u7cd6\u57fa\u5316\u6216\u5206\u6ccc\u7684\uff0c\u60a8\u53ef\u80fd\u5e0c\u671b\u9078\u64c7pYLSC1\u8f09\u9ad4\u3002<\/p>\n

pYLEX1\u8868\u9054\u8f09\u9ad4\uff087259bp\uff09\u542b\u6709\u5f37\u96dc\u5408\u555f\u52d5\u5b50\uff08hp4d\uff09\uff0c\u5176\u651c\u5e36\u4f86\u81eapXPR2\u7684\u56db\u500b\u4e32\u806f\u62f7\u8c9d\u7684\u4e0a\u6e38\u6fc0\u6d3b\u5b50\u5e8f\u5217\uff08UAS1B\uff09\u7247\u6bb5\u548c\u6700\u5c0f\u7684pLEU2\u7247\u6bb5\u3002\u591a\u514b\u9686\u4f4d\u9ede\u548cXPR2\u8f49\u9304\u7d42\u6b62\u5b50\u7dca\u9130hp4d\u555f\u52d5\u5b50\u76843’\u4f4d\u9ede\u7684\u4e0b\u6e38\uff0c\u7136\u5f8c\u662f\u4eae\u6c28\u9178\u9078\u64c7\u6a19\u8a18\u57fa\u56e0\uff08LEU2\uff09\u3002\u901a\u904e\u7528NotI\u6d88\u5316\u53ef\u4ee5\u4f7f\u8f09\u9ad4\u7dda\u6027\u5316\uff0c\u4ee5\u7522\u751f\u80fd\u5920\u63d2\u5165\u89e3\u8102\u8036\u6c0f\u9175\u6bcd\u57fa\u56e0\u7d44\u7684\u7dda\u6027DNA\u7247\u6bb5\u3002<\/p>\n

pYLSC1\u5206\u6ccc\u8f09\u9ad4\uff087205bp\uff09\u542b\u6709\u96dc\u5408\u555f\u52d5\u5b50\uff08hp4d\uff09\u548c\u5206\u6ccc\u4fe1\u865f\uff08XPR2\u524d\u5340\uff09\u3002\u591a\u514b\u9686\u4f4d\u9ede\u548cpXPR2\u8f49\u9304\u7d42\u6b62\u5b50\u7dca\u9130XPR2\u524d\u5340\u76843’\u4f4d\u9ede\u7684\u4e0b\u6e38\uff0c\u63a5\u8457\u662f\u4eae\u6c28\u9178\u9078\u64c7\u6a19\u8a18\u57fa\u56e0\uff08LEU2\uff09\u3002\u901a\u904e\u7528NotI\u6d88\u5316\u53ef\u4ee5\u4f7f\u8f09\u9ad4\u7dda\u6027\u5316\uff0c\u4ee5\u7522\u751f\u80fd\u5920\u63d2\u5165\u89e3\u8102\u8036\u6c0f\u9175\u6bcd\u57fa\u56e0\u7d44\u7684\u7dda\u6027DNA\u7247\u6bb5\u3002
\u9175\u6bcd\u83cc\u682a
\u89e3\u8102\u8036\u6c0f\u9175\u6bcd\u7684\u83cc\u682aPo1g\u662f\u91ce\u751f\u578b\u83cc\u682aW29\uff08ATCC 20460\uff09\u7684\u884d\u751f\u7269\uff0c\u901a\u904e\u4e00\u7cfb\u5217\u907a\u50b3\u4fee\u98fe\u3002\u7c21\u8a00\u4e4b\uff0c\u7528\u4f86\u81ea\u91c0\u9152\u9175\u6bcd\u7684SUC2\u57fa\u56e0\u7834\u58deW29\u83cc\u682a\u4e2d\u7684\u539f\u59cbURA3\u57fa\u56e0\uff0c\u7136\u5f8c\u5728LEU2\u57fa\u56e0\u4e2d\u5f15\u5165\u7f3a\u5931\u3002\u6b64\u5916\uff0cXPR2\u548cAXP\u57fa\u56e0\u7684\u7f3a\u5931\u78ba\u4fddPo1g\u4e0d\u80fd\u7522\u751f\u4efb\u4f55\u7d30\u80de\u5916\u86cb\u767d\u9176\u3002\u70ba\u4e86\u5bb9\u6613\u5730\u6574\u5408\u57fa\u65bcpBR\u7684\u8868\u9054\/\u5206\u6ccc\u8f09\u9ad4\uff0c\u5c07pBR322\u5c0d\u63a5\u5e73\u53f0\u6574\u5408\u5230URA3\u57fa\u56e0\u5ea7\u3002<\/p>\n\n\n\n\n
Strain<\/strong><\/td>\nGenotype<\/strong><\/td>\nPhenotype<\/strong><\/td>\n<\/tr>\n
Po 1 g<\/td>\n\n

Mat<\/em>A,\u00a0leu<\/em>2-270,\u00a0ura<\/em>3-302::URA3,
xpr<\/em>2-332,\u00a0axp<\/em>-2<\/p>\n<\/td>\n

Leu-, \u0394AEP, \u0394AXP, Suc+, pBR platform<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

\u8cea\u91cf\u63a7\u5236<\/strong>
\u4f7f\u7528\u5169\u7a2e\u8cea\u7c92pYLEX1 + AMY1\uff08\u5c0f\u9f20\u553e\u6db2\u03b1-\u6fb1\u7c89\u9176\u57fa\u56e0\uff09\u548cpYLSC1 +AMY1\u0394\uff08\u6c92\u6709\u5176\u5929\u7136\u5206\u6ccc\u4fe1\u865f\u7684AMY1\uff09\u4f86\u78ba\u4fdd\u529f\u80fd\u6027\u8f49\u578b\u8a66\u5291\u76d2\u548c\u5bbf\u4e3b\u4e2d\u6b63\u78ba\u7684\u57fa\u56e0\u8868\u9054\u3002\u9084\u9032\u884c\u6c61\u67d3\u6e2c\u8a66\u4ee5\u78ba\u4fdd\u6c92\u6709\u5176\u4ed6\u5fae\u751f\u7269\u6c61\u67d3\u3002
\u5be6\u9a57\u6578\u64da
\u8a72\u5716\u986f\u793a\u4f86\u81ea\u7de8\u78bc\u6fb1\u7c89\u9176\u7684\u8f49\u5316\u9ad4\uff08YL\uff032\u548c\uff033\uff09\u7684\u5206\u6279\u57f9\u990a\u7684\u904e\u6ffe\u57f9\u990a\u57fa\u53ef\u4ee5\u5728\u56fa\u9ad4\u57f9\u990a\u57fa\u74ca\u8102\u4e2d\u6d88\u5316\u6fb1\u7c89\uff0c\u4e26\u96a8\u5f8c\u7522\u751f\u900f\u660e\u5340\u57df\u3002\u76f8\u53cd\uff0c\u4f86\u81ea\u50c5\u7528\u8f09\u9ad4\u8f49\u5316\u7684\u9175\u6bcd\u57f9\u990a\u57fa\uff08YL\uff031\uff09\u7684\u57f9\u990a\u57fa\u6c92\u6709\u8868\u73fe\u51fa\u76f8\u540c\u7684\u7d50\u679c\u3002\u9019\u8868\u660e\u901a\u904e\u4f7f\u7528YLEX\u8868\u9054\u8a66\u5291\u76d2\u6210\u529f\u5730\u5c07\u03b1-\u6fb1\u7c89\u9176\u57fa\u56e0\uff08AMY1\uff09\u514b\u9686\u5230pYLEX1\u548c\u03b1-\u6fb1\u7c89\u9176\u57fa\u56e0\u800c\u6c92\u6709\u5176\u5206\u6ccc\u4fe1\u865f\u80bd\uff08AMY1D\uff09\u5230pYLSC1\u4e2d\u3002\u5728\u5169\u7a2e\u60c5\u6cc1\u4e0b\uff0c\u6d3b\u6027\u03b1-\u6fb1\u7c89\u9176\u6709\u6548\u5730\u5206\u6ccc\u5230\u57f9\u990a\u57fa\u4e2d\u3002<\/p>\n

YLEX\u7cfb\u7d71\uff08\u89e3\u8102\u8036\u6c0f\u9175\u6bcd\u8868\u9054\u7cfb\u7d71\uff09\u4e2d\u9ad8\u7d30\u80de\u5bc6\u5ea6\u767c\u9175\u548c\u91cd\u7d44\u9176\u5206\u6ccc\u80fd\u529b\u7684\u6642\u7a0b\u7814\u7a76\u3002<\/strong><\/p>\n

\"\"<\/p>\n

\u7570\u6e90\u86cb\u767d\u8cea\u7684\u904e\u91cf\u7522\u751f\uff08SDS-PAGE\u5206\u6790\uff09\uff1a\u4e0a\u6e05\u6db2\u4e2d\u91cd\u7d44\u9176\u7684\u5206\u6ccc\u8b5c\uff0c\u901a\u904e\u6642\u9593\u904e\u7a0b\u6536\u96c6\u6a23\u54c1\uff0c10\u03bcL\/\u6cf3\u9053\u3002<\/p>\n

\"\"<\/p>\n

\u9ad8\u7d30\u80de\u5bc6\u5ea6\u767c\u9175\uff1a\u572814L\u88dc\u6599\u5206\u6279\u767c\u9175\u7f50\u4e2d\u9032\u884c\u9ad8\u7d30\u80de\u5bc6\u5ea6\u767c\u9175\uff0c\u901a\u904e\u6642\u9593\u904e\u7a0b\u96e2\u5fc3\uff083,000rpm\uff09\u7d30\u80de\u3002<\/p>\n

\"\"<\/p>\n

* YLEX Expression Kit\uff08\u7522\u54c1\u898f\u683c\uff09
\u89e3\u8102\u8036\u6c0f\u9175\u6bcd\u83cc\u682a\uff1aPo1g; pYLEX1-\u8868\u9054\u8f09\u9ad41\u523a; pYLSC1-\u5206\u6ccc\u8f09\u9ad45\u03bcg;\u5f15\u72696560F\uff08\u7528\u65bc\u6e2c\u5e8f\u76ee\u7684\uff09250\u03bcL;\u5f15\u72696904F\uff08\u7528\u65bc\u6e2c\u5e8f\u76ee\u7684\uff09250\u03bcL; YLOS\u8f49\u5316\u8a66\u5291\u76d21\u8a66\u5291\u76d2\uff08120 rxns\uff09\u3002<\/p>\n

* YLEX Expression Kit\u7531Yeastern Biotech Co.\uff0cLtd\u3002\u751f\u7522\uff0c\u5c08\u5229\u4fdd\u8b77\uff0c\u7531\u6cd5\u570b\u570b\u5bb6\u8fb2\u696d\u7814\u7a76\u6240\uff08INRA\uff09\u7684\u5c08\u6709\u8ca1\u7522\u6388\u6b0a\uff0c\u5728\u5168\u7403\u7bc4\u570d\u5167\u9032\u884c\u975e\u7368\u5bb6\u88fd\u9020\u548c\u5206\u92b7\u3002<\/p>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n

\n
\n
\n
\n

\u898f\u7bc4
\u5b58\u653e\u5728<\/p>\n

\"\"<\/p>\n

\n

# FYY201-1KT<\/b><\/p>\n

\n\n\n\n\n\n\n\n\n
Yarrowia lipolytica<\/em>\u00a0Yeast Strain: Po1g<\/td>\n1 stab<\/td>\n<\/tr>\n
pYLEX1<\/em>\u00a0\u2013 Expression Vector<\/td>\n5 \u03bcg<\/td>\n<\/tr>\n
pYLSC1<\/em>\u00a0\u2013 Secretion Vector<\/td>\n5\u00a0\u03bcg<\/td>\n<\/tr>\n
Primer 6560 F<\/td>\n250\u00a0\u03bcl<\/td>\n<\/tr>\n
Primer 6904 R<\/td>\n250\u00a0\u03bcl<\/td>\n<\/tr>\n
YLOS<\/em>\u00a0Transformation Kit
(# FYY301-120P)<\/strong><\/td>\n		1 kit<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<\/div>\n<\/div>\n\n\n\n
\n

Manual<\/p>\n

\"\"<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n","protected":false},"excerpt":{"rendered":"

YLEX\u9175\u6bcd\u8868\u9054\u8a66\u5291\u76d2\u63cf\u8ff0\u57fa\u65bcINRA INAPG\u8a31\u53ef\u5c08\u5229*\u7684YLEX Expression Kit\u63d0\u4f9b\u4e86\u4e00\u7a2e […]<\/p>\n","protected":false},"author":20,"featured_media":0,"parent":0,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"","meta":{"_et_pb_use_builder":"","_et_pb_old_content":"","_et_gb_content_width":"","footnotes":""},"_links":{"self":[{"href":"https:\/\/si.secda.info\/yb_mdn\/index.php?rest_route=\/wp\/v2\/pages\/3191"}],"collection":[{"href":"https:\/\/si.secda.info\/yb_mdn\/index.php?rest_route=\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/si.secda.info\/yb_mdn\/index.php?rest_route=\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/si.secda.info\/yb_mdn\/index.php?rest_route=\/wp\/v2\/users\/20"}],"replies":[{"embeddable":true,"href":"https:\/\/si.secda.info\/yb_mdn\/index.php?rest_route=%2Fwp%2Fv2%2Fcomments&post=3191"}],"version-history":[{"count":3,"href":"https:\/\/si.secda.info\/yb_mdn\/index.php?rest_route=\/wp\/v2\/pages\/3191\/revisions"}],"predecessor-version":[{"id":3201,"href":"https:\/\/si.secda.info\/yb_mdn\/index.php?rest_route=\/wp\/v2\/pages\/3191\/revisions\/3201"}],"wp:attachment":[{"href":"https:\/\/si.secda.info\/yb_mdn\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=3191"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}