1.1 \u6750\u6599<\/strong><\/p>\n\n\n\n 1.1.1 \u83cc\u682a\u548c\u8d28\u7c92<\/strong><\/p>\n\n\n\n \u672c\u7814\u7a76\u6240\u7528\u521d\u59cb\u83cc\u682a\u4e0e\u8d28\u7c92\u5747\u4e3a\u5b9e\u9a8c\u5ba4\u4fdd\u85cf\u3002\u8be6\u7ec6\u4fe1\u606f\u89c1\u8868 1 \u548c\u8868 2\u3002<\/p>\n\n\n\n 1.1.2 \u57f9\u517b\u57fa<\/strong><\/p>\n\n\n\n LB \u57f9\u517b\u57fa\uff1a5 g\/L \u9175\u6bcd\u7c89\uff0c10 g\/L \u86cb\u767d\u80e8\uff0c10 g\/L Na Cl\uff0c\u56fa\u4f53\u57f9\u517b\u57fa\u6dfb\u52a0 20 g\/L \u7684\u743c\u8102\u7c89\u3002\u6839\u636e\u9700\u8981\u6dfb\u52a0\u76f8\u5e94\u6d53\u5ea6\u7684\u6297\u751f\u7d20(\u6c2f\u9709\u7d20\u7ec8\u6d53\u5ea6\u4e3a 5 mg\/L\uff0c\u58ee\u89c2\u9709\u7d20\u7ec8\u6d53\u5ea6\u4e3a 100 mg\/L\uff0c\u6c28\u82c4\u9709\u7d20\u7ec8\u6d53\u5ea6\u4e3a 100 mg\/L)\u3002<\/p>\n\n\n\n 1.1.3 \u4e3b\u8981\u8bd5\u5242<\/strong><\/p>\n\n\n\n Prime Star max \u805a\u5408\u9176\u8d2d\u4e8e Ta Ka Ra \u516c\u53f8\uff1b\u8d28\u7c92\u63d0\u53d6\u3001\u611f\u53d7\u6001\u5236\u5907\u8bd5\u5242\u76d2\u8d2d\u4e8e\u751f\u5de5\u751f\u7269\u5de5\u7a0b(\u4e0a\u6d77)\u80a1\u4efd\u6709\u9650\u516c\u53f8\uff1bGene JET PCR \u7eaf\u5316\u8bd5\u5242\u76d2\u8d2d\u4e8e Thermo Scientific \u516c\u53f8\u3002<\/p>\n\n\n\n 1.2 \u65b9\u6cd5<\/strong> \u57fa\u56e0\u5e8f\u5217\u4e0e\u5f15\u7269\u5408\u6210<\/strong><\/p>\n\n\n\n \u5c06 NCBI \u6570\u636e\u5e93\u4e2d B. breve UCC2003 \u6765\u6e90Nan R \u7684\u6c28\u57fa\u9178\u5e8f\u5217\u9001\u81f3\u82cf\u5dde\u91d1\u552f\u667a\u516c\u53f8\u8fdb\u884c\u5bc6\u7801\u5b50\u4f18\u5316(\u8868\u8fbe\u5bbf\u4e3b\uff1aBacillus subtilis 168)\u5e76\u5408\u6210\uff0cBbr_Nan R \u7ed3\u5408\u4f4d\u70b9\u53c2\u8003 Egan \u7b49[21]\u7684\u62a5\u9053\u3002\u6587\u4e2d\u6240\u6709\u5f15\u7269\u7684\u5408\u6210\u548c\u6d4b\u5e8f\u53cd\u5e94\u5747\u7531\u82cf\u5dde\u91d1\u552f\u667a\u751f\u7269\u79d1\u6280\u6709\u9650\u516c\u53f8\u6216\u8005\u751f\u5de5\u751f\u7269\u5de5\u7a0b(\u4e0a\u6d77)\u80a1\u4efd\u6709\u9650\u516c\u53f8(\u4ee5\u4e0b\u7b80\u79f0\u201c\u4e0a\u6d77\u751f\u5de5\u201d)\u5b8c\u6210\uff0c\u672c\u6587\u90e8\u5206\u5f15\u7269\u4fe1\u606f\u89c1\u8868 3\uff0c\u66f4\u6362\u542f\u52a8\u5b50\u6240\u7528\u5f15\u7269\u6839\u636e\u8868 4 \u542f\u52a8\u5b50\u5e8f\u5217\u8fdb\u884c\u8bbe\u8ba1\u3002<\/p>\n\n\n\n 1.2.2 \u91cd\u7ec4\u83cc\u682a\u7684\u6784\u5efa<\/strong><\/p>\n\n\n\n BS-nan C\u3001BS-nan T \u7684\u6784\u5efa\uff1a\u4ee5 B. subtilis 168\u57fa\u56e0\u7ec4\u4e3a\u6a21\u677f\uff0c\u901a\u8fc7\u805a\u5408\u9176\u94fe\u5f0f\u53cd\u5e94(polymerasechain reaction, PCR)\u6269\u589e ydb D \u4f4d\u70b9\u4e0a\u4e0b\u6e38\u54041 000 bp \u7684\u5e8f\u5217\u4f5c\u4e3a\u5de6\u53f3\u540c\u6e90\u81c2\uff0c\u4ee5 E. coli JM109 \u57fa\u56e0\u7ec4\u4e3a\u6a21\u677f\uff0c\u6269\u589e nan C\u3001nan T \u57fa\u56e0\uff0c\u4ee5\u5b9e\u9a8c\u5ba4\u4fdd\u5b58\u8d28\u7c92 p7S6P43[7]\u4e3a\u6a21\u677f\u6269\u589e\u58ee\u89c2 \u9709\u7d20\u6297\u6027\u6846\u548c P43 \u542f\u52a8\u5b50\u7247\u6bb5\u3002\u901a\u8fc7\u878d\u5408 PCR\u5c06\u5de6\u53f3\u81c2\u3001\u6297\u6027\u6846\u548c P43 \u542f\u52a8\u5b50\u7247\u6bb5\u3001nan C (nan T)\u7247\u6bb5\u8fdb\u884c\u878d\u5408\uff0c\u83b7\u5f97\u57fa\u56e0 nan C (nan T)\u7684\u8868\u8fbe\u6846\uff0c\u5e76\u8f6c\u5316\u67af\u8349\u82bd\u5b62\u6746\u83cc BSXC \u611f\u53d7\u6001\uff0c\u83b7\u5f97\u83cc\u682a BS-nan C (BS-nan T)\u3002 BSP1\u2013BSP10 \u7684\u6784\u5efa\uff1a\u5de6\u53f3\u81c2\u548c nan T \u7247\u6bb5\u53c2\u8003\u4e0a\u8ff0\u65b9\u6cd5\u83b7\u5f97\uff0c\u4ee5\u5b9e\u9a8c\u5ba4\u4fdd\u5b58\u7684 p7S6P1<\/p>\n\n\n\n \u7cfb\u5217\u8d28\u7c92[22]\u4e3a\u6a21\u677f\u6269\u589e\u83b7\u5f97\u58ee\u89c2\u9709\u7d20\u6297\u6027\u6846\u548c\u542f\u52a8\u5b50\u7247\u6bb5\uff0c\u7136\u540e\u6309\u7167\u4e0a\u8ff0 BS-nan T \u7684\u6784\u5efa\u65b9\u6cd5\u5f97\u5230\u91cd\u7ec4\u83cc\u682a BSP1\u2013BSP10\u3002<\/p>\n\n\n\n 1.2.3 \u91cd\u7ec4\u8d28\u7c92\u7684\u6784\u5efa<\/strong><\/p>\n\n\n\n \u6742 \u5408 \u542f \u52a8 \u5b50 \u91cd \u7ec4 \u8d28 \u7c92 \u7684 \u6784 \u5efa \uff1a \u4ee5 p HT- veg-gfp \u4e3a\u6a21\u677f\uff0c\u7528\u76f8\u5e94\u7684\u5f15\u7269\u8fdb\u884c\u6269\u589e\u83b7\u5f97\u5e26\u6709\u6742\u5408\u542f\u52a8\u5b50\u7684\u7ebf\u6027\u8d28\u7c92\u7247\u6bb5\uff0c\u4e0a\u4e0b\u6e38\u5f15\u7269\u5305\u62ec\u6742\u5408\u542f\u52a8\u5b50\u5e8f\u5217\u3001\u7279\u5f02\u6027\u7ed3\u5408\u5e8f\u5217\uff0c\u4e14\u5b58\u5728\u4e00\u6bb5 20 bp \u5de6\u53f3\u7684\u91cd\u53e0(overlap)\u3002\u5229\u7528 Thermo Scientific Gene JET PCR \u7eaf\u5316\u8bd5\u5242\u76d2\u5bf9\u6240\u83b7\u5f97\u7684\u7ebf\u6027\u8d28\u7c92\u7247\u6bb5\u8fdb\u884c\u7eaf\u5316\uff0c\u7136\u540e\u53d6 10 \u03bcL \u8f6c\u5316E. coli JM109 \u611f\u53d7\u6001\uff0c\u5e76\u6d82\u5e03\u6297\u6027\u5e73\u677f\u3002\u6b21\u65e5\u5c06\u5e73\u677f\u4e0a\u7684\u5355\u83cc\u843d\u9001\u4e0a\u6d77\u751f\u5de5\u6d4b\u5e8f\uff0c\u5c06\u6d4b\u5e8f\u6b63\u786e\u7684\u5355\u83cc\u843d\u6269\u5927\u57f9\u517b\uff0c\u5e76\u63d0\u53d6\u91cd\u7ec4\u8d28\u7c92\u3002\u4f18\u5316nan R \u8868\u8fbe\u91cf\u65f6\uff0c\u4e5f\u91c7\u7528\u540c\u6837\u7684\u73af\u5316 PCR \u7684\u65b9\u5f0f\u66ff\u6362 nan R \u7684\u542f\u52a8\u5b50\u3002<\/p>\n\n\n\n \u751f\u7269\u4f20\u611f\u5668\u91cd\u7ec4\u8d28\u7c92\u7684\u6784\u5efa\uff1a\u5206\u522b\u4ee5\u6742\u5408\u542f\u52a8\u5b50\u91cd\u7ec4\u8d28\u7c92\u4e3a\u6a21\u677f\uff0c\u7528\u5f15\u7269 p N-F\/R \u6269\u589e\u8d28\u7c92\u9aa8\u67b6\u53ca\u6742\u5408\u542f\u52a8\u5b50\u8868\u8fbe\u7684\u7eff\u8272\u8367\u5149\u86cb\u767d\u57fa\u56e0gfp\uff0c\u4ee5 p HT-nan R \u4e3a\u6a21\u677f\uff0c\u7528\u5f15\u7269 nan R-F\/R \u6269\u589e Prps T \u542f\u52a8\u5b50\u8868\u8fbe\u7684 nan R \u57fa\u56e0\uff0c\u5229\u7528 Thermo Scientific Gene JET PCR \u7eaf\u5316\u8bd5\u5242\u76d2\u5c06\u6240\u5f97\u7247\u6bb5\u7eaf\u5316\uff0c\u7ecf Gibson \u7ec4\u88c5[23]\u540e\u8f6c\u5316 E. coli JM109\u611f\u53d7\u6001\uff0c\u5e76\u6d82\u5e03\u6297\u6027\u5e73\u677f\u3002\u6b21\u65e5\u5c06\u5e73\u677f\u4e0a\u7684\u5355\u83cc\u843d\u9001\u4e0a\u6d77\u751f\u5de5\u6d4b\u5e8f\uff0c\u5c06\u6d4b\u5e8f\u6b63\u786e\u7684\u5355\u83cc\u843d\u6269\u5927\u57f9\u517b\uff0c\u5e76\u63d0\u53d6\u91cd\u7ec4\u8d28\u7c92\u3002<\/p>\n\n\n\n 1.2.4 \u611f\u53d7\u6001\u7ec6\u80de\u7684\u5236\u5907\u548c\u8f6c\u5316<\/strong><\/p>\n\n\n\n \u5927\u80a0\u6746\u83cc\u611f\u53d7\u6001\u5236\u5907\u4e0e\u8f6c\u5316\uff1a\u8d85\u7ea7\u611f\u53d7\u6001\u7ec6\u80de\u5236\u5907\u8bd5\u5242\u76d2\u8d2d\u4e8e\u751f\u5de5\u751f\u7269\u5de5\u7a0b(\u4e0a\u6d77)\u80a1\u4efd\u6709\u9650\u516c\u53f8\uff0c\u6309\u7167\u8bf4\u660e\u4e66\u8981\u6c42\u5236\u5907\u3001\u8f6c\u5316E. coli JM109 \u611f\u53d7\u6001\u7ec6\u80de\uff0c\u7528\u4e8e\u8d28\u7c92\u7684\u6784\u5efa\u3002 \u67af\u8349\u82bd\u5b62\u6746\u83cc\u611f\u53d7\u6001\u5236\u5907\u4e0e\u8f6c\u5316\uff1a\u53c2\u8003Zhang \u7b49[24]\u7684\u65b9\u6cd5\uff0c\u5728 B. subtilis 168 \u57fa\u56e0\u7ec4\u4e0a\u6574\u5408\u4e86\u4e00\u4e2a\u53d7\u6728\u7cd6\u8bf1\u5bfc\u578b\u542f\u52a8\u5b50Pxyl A\u8c03\u63a7\u7684com K\u57fa\u56e0\uff0c\u5f97\u5230\u83cc\u682aBSXC\u4f5c\u4e3a\u672c\u7814\u7a76\u7684\u51fa\u53d1\u83cc\u682a\uff0c\u5229\u7528\u6728\u7cd6\u8bf1\u5bfc\u5236\u5907\u611f\u53d7\u6001\u7ec6\u80de\u3002\u6311\u53d6\u5e73\u677f\u6d3b\u5316\u7684B. subtilis \u5355\u83cc\u843d\u57281mL LB \u57f9\u517b\u57fa\u4e2d\u4e8e 37\u2103\u3001220 r\/min \u7684\u6761\u4ef6\u4e0b\u57f9\u517b 12h\uff0c\u52a0\u5165LB \u57f9\u517b\u57fa\u7a00\u91ca\u81f35mL\uff0c\u5e76\u52a0\u5165\u7ec8\u6d53\u5ea6\u4e3a 3%\u7684\u6728\u7cd6\uff0c\u4e8e 37\u2103\u3001220 r\/min \u6761\u4ef6\u4e0b\u8bf1\u5bfc 2 h \u540e\u83b7\u5f97\u611f\u53d7\u6001\u3002\u5c06\u9002\u91cf\u7684\u8d28\u7c92\u6216\u7247\u6bb5\u52a0\u5165100 \u00b5LB. subtilis \u611f\u53d7\u6001\u7ec6\u80de\uff0c\u4e8e37\u2103\u3001220 r\/min\u7684\u6761\u4ef6\u4e0b\u57f9\u517b 1.5 h\u540e\u6d82\u5e03\u76f8\u5e94\u6297\u6027\u5e73\u677f\uff0c\u7f6e\u4e8e37\u2103\u6052\u6e29\u57f9\u517b\u7bb1\u57f9\u517b\u3002<\/p>\n\n\n\n 1.2.5 \u80de\u5185 Neu Ac \u6d53\u5ea6\u68c0\u6d4b<\/strong><\/p>\n\n\n\n \u80de\u5185 Neu Ac \u6d4b\u5b9a\u6837\u54c1\u5236\u5907\uff1a\u53d61 m L\u53d1\u9175\u6db2\uff0c\u7f6e\u4e8e\u9884\u51b7\u4e3a4\u2103\u7684\u79bb\u5fc3\u673a\uff0c6 000 r\/min \u79bb\u5fc3 5 min\uff0c\u5f03\u5c3d\u4e0a\u6e05\u6db2\uff0c\u5c06\u83cc\u4f53\u91cd\u60ac\u4e8e 1 m L \u9884\u51b7\u7684\u65e0\u83cc\u6c34\u4e2d\u6d17\u6da41\u6b21\uff0c6 000 r\/min \u79bb\u5fc3 5 min\uff0c\u5f03\u5c3d\u4e0a\u6e05\u6db2\uff0c\u7528 0.2 m L \u65e0\u83cc\u6c34\u91cd\u60ac\u83cc\u4f53\uff0c\u540e\u52a0\u51650.4 m L\u4e59\u8148\u548c 0.4 m L\u7532\u9187\u6df7\u5300\uff0c\u5e76\u7f6e\u4e8e\u201320 \u2103\u8fc7\u591c\u8403\u53d6\u3002\u5c06\u8403\u53d6\u6db2\u4e8e4\u2103\u300112 000 r\/min \u79bb\u5fc310 min\uff0c\u4e0a\u6e05\u6db2\u7f6e\u4e8e\u51b7\u51bb\u5e72\u71e5\u673a\u8fc7\u591c\u5e72\u71e5\u3002\u5e72\u71e5\u540e\u7684\u7c89\u672b\u91cd\u60ac\u4e8e\u4e00\u5b9a\u4f53\u79ef\u7684\u65e0\u83cc\u6c34\u4ee5\u6d53\u7f29\u6837\u54c1\uff0c\u7528\u4e8e\u9ad8\u6548\u6db2\u76f8\u8272\u8c31\u68c0\u6d4b\u3002 \u9ad8\u6548\u6db2\u76f8\u8272\u8c31(high performance liquid chromatography, HPLC) \u68c0\u6d4b\uff1a\u53d6\u6837\u54c1\u901a\u8fc70.22\u03bcm \u7684\u6ee4\u819c\uff0c\u91c7\u7528 Aminex HPX-87H \u67f1 (300 nm\u00d77.8 nm)\u8fdb\u884c HPLC \u68c0\u6d4b\uff0c\u68c0\u6d4b\u53c2\u6570\u5982\u4e0b\uff1a\u91c7\u752810 mmol\/L H2SO4\u4f5c\u4e3a\u6d41\u52a8\u76f8\uff0c0.5 m L\/min\u6d41\u901f\u8fdb\u6837\uff0c\u8fdb\u6837\u91cf\u4e3a10\u03bcL\uff0c\u67f1\u6e29\u4e3a40 \u2103\uff0c\u7d2b\u5916\u68c0\u6d4b\u5668\u6ce2\u957f\u4e3a210 nm\u3002<\/p>\n\n\n\n 1.2.6 GFP \u8367\u5149\u5f3a\u5ea6\u68c0\u6d4b\u65b9\u6cd5<\/strong><\/p>\n\n\n\n \u5c06\u57f9\u517b\u8fc7\u591c\u7684\u79cd\u5b50\u6db2\u4ee5 2%\u8f6c\u63a5\u5230\u542b\u6709 0.2 mL LB \u57f9\u517b\u57fa\u7684 96 \u6d45\u5b54\u677f\u4e2d\uff0c37 \u2103\u3001900 r\/min \u57f9\u517b 8h\uff0c\u7528\u9176\u6807\u4eea\u5728490nm \u7684\u6fc0\u53d1\u6ce2\u957f\u3001530nm\u7684\u53d1\u5c04\u6ce2\u957f\u548c 60 \u7684\u589e\u76ca\u503c\u4e0b\u6d4b\u5b9a GFP\u7684\u8367\u5149\u5f3a\u5ea6\uff1b\u5728 600nm \u7684\u6ce2\u957f\u4e0b\u6d4b\u5b9a OD600\u3002\u51cf\u53bb\u57f9\u517b\u57fa\u7684\u80cc\u666f\u566a\u58f0\u540e\uff0c\u8ba1\u7b97\u8367\u5149\u5f3a\u5ea6\u4e0eOD600\u7684\u6bd4\u503c\uff0c\u518d\u51cf\u53bb\u65e0GFP \u8868\u8fbe\u7684\u5bf9\u7167\u83cc\u682a\u7684\u8367\u5149\u5f3a \u5ea6\u4e0eOD600\u7684\u6bd4\u503c\uff0c\u5373\u5f97\u5230\u76f8\u5bf9\u8367\u5149\u5f3a\u5ea6\u3002<\/p>\n\n\n\n 2 \u7ed3\u679c\u4e0e\u5206\u6790<\/strong><\/p>\n\n\n\n 2.1 Neu Ac \u8f6c\u8fd0\u86cb\u767d Nan T \u548c Nan C \u7684\u6d4b\u8bd5\u548c\u8868\u8fbe\u4f18\u5316<\/strong><\/p>\n\n\n\n \u5728\u54cd\u5e94\u5c0f\u5206\u5b50\u4ee3\u8c22\u7269\u7684\u751f\u7269\u4f20\u611f\u5668\u6784\u5efa\u8fc7\u7a0b\u4e2d\uff0c\u5e38\u5e38\u91c7\u7528\u5728\u57f9\u517b\u57fa\u4e2d\u6dfb\u52a0\u4e0d\u540c\u6d53\u5ea6\u7684\u5c0f\u5206\u5b50\u4ee3\u8c22\u7269\u6765\u9a8c\u8bc1\u751f\u7269\u4f20\u611f\u5668\u7684\u54cd\u5e94\u6027\u80fd\uff0c\u800c\u67af\u8349\u82bd\u5b62\u6746\u83cc\u4e0d\u5177\u5907\u5c06\u80de\u5916 Neu Ac \u8f6c\u8fd0\u8fdb\u80de\u5185\u7684\u80fd\u529b\uff0c\u56e0\u6b64\uff0c\u9996\u5148\u9700\u8981\u6784\u5efa\u80fd\u5c06\u80de\u5916 Neu Ac\u8f6c\u8fd0\u8fdb\u80de\u5185\u7684\u67af\u8349\u82bd\u5b62\u6746\u83cc\uff0c\u4fbf\u4e8e\u5916\u6e90\u6dfb\u52a0Neu Ac \u4ee5\u9a8c\u8bc1\u751f\u7269\u4f20\u611f\u5668\u7684\u6027\u80fd\u3002\u4e3a\u4e86\u4e0d\u5f71\u54cd\u5185\u6e90\u57fa\u56e0\u7684\u8868\u8fbe\uff0c\u9009\u62e9 ydb C \u548c ydb D \u57fa\u56e0\u7684\u95f4\u9694\u4f4d\u7f6e\u4f5c\u4e3a\u57fa\u56e0\u7ec4\u6574\u5408\u8868\u8fbe nan C \u548c nan T \u7684\u4f4d\u70b9\u3002\u6309\u7167\u65b9\u6cd5 1.1.2\uff0c\u5c06\u7531 P43 \u542f\u52a8\u5b50\u8c03\u63a7 nan C (nan T)\u7684\u8868\u8fbe\u6846\u6574\u5408\u5230\u67af\u8349\u82bd\u5b62\u6746\u83cc BSXC \u7684\u57fa\u56e0\u7ec4\uff0c\u5f97\u5230\u83cc\u682a BS-nan C (BS-nan T)\u3002\u4e3a\u4e86\u6bd4\u8f83 Nan C \u548c Nan T \u8f6c\u8fd0 Neu Ac \u7684\u80fd\u529b\uff0c\u5c06\u83cc\u682a BS-nan C \u548c BS-nan T \u5206\u522b\u5728\u6dfb\u52a0\u4e86 10g\/L\u548c 80g\/L Neu Ac \u7684 LB \u57f9\u517b\u57fa\u4e2d\u53d1\u9175 12h\uff0c\u968f\u540e\u68c0\u6d4b\u5176\u80de\u5185 Neu Ac \u542b\u91cf\u3002<\/p>\n\n\n\n \u4e0e BS-nan C \u76f8\u6bd4\uff0cBS-nan T \u8f6c\u8fd0 Neu Ac \u7684\u80fd\u529b\u66f4\u5f3a\uff0c\u4e14\u5916\u6e90\u6dfb\u52a0\u4e0d\u540c\u6d53\u5ea6\u7684 Neu Ac \u65f6\u80de\u5185\u6d53\u5ea6\u5dee\u5f02\u8f83\u5927\uff0c\u662f\u9a8c\u8bc1\u54cd\u5e94 Neu Ac \u7684\u751f\u7269\u4f20\u611f\u5668\u7684\u66f4\u4f18\u5bbf\u4e3b(\u56fe 1A)\u3002BS-nan T \u5728\u5916\u6e90\u6dfb\u52a010\u201380 g\/L Neu Ac \u7684 LB \u57f9\u517b\u57fa\u4e2d\u53d1\u9175 12 h \u65f6\uff0c\u80de\u5185 Neu Ac \u6d53\u5ea6\u4e5f\u5b58\u5728\u5dee\u5f02\uff0c\u4f46\u662f\u80de\u5185 Neu Ac\u6d53\u5ea6\u603b\u4f53\u504f\u4f4e\uff0c\u6700\u9ad8\u4e5f\u53ea\u6709 310 \u03bcg\/m L (\u56fe 1B)\uff0c\u8fd9\u4e0d\u5229\u4e8e\u5bf9\u54cd\u5e94 Neu Ac \u6d53\u5ea6\u8303\u56f4\u5e7f\u7684\u751f\u7269\u4f20\u611f\u5668\u7684\u9a8c\u8bc1\uff0c\u4e8e\u662f\u8fdb\u4e00\u6b65\u5bf9 nan T \u8fdb\u884c\u8868\u8fbe\u4f18\u5316\u3002\u9009\u53d6\u67af\u8349\u82bd\u5b62\u6746\u83cc\u4e2d 9 \u4e2a\u4e0d\u540c\u5f3a\u5ea6\u7684\u5185\u6e90\u542f\u52a8\u5b50(P1\u3001P2\u3001P3\u3001P4\u3001P6\u3001P7\u3001P8\u3001P9 \u548c P10)[22]\u66ff\u6362 P43 \u542f\u52a8\u5b50\u8c03\u63a7 nan T \u7684\u8868\u8fbe\uff0c\u5f97\u5230\u83cc\u682aBSP1\u2013BSP10\u3002\u5982\u56fe 2 \u6240\u793a\uff0c\u5f53\u5728\u6dfb\u52a0\u4e86 10 g\/L Neu Ac \u7684 LB \u57f9\u517b\u57fa\u4e2d\u53d1\u9175 12 h \u65f6\uff0c\u83cc\u682aBSP1\u2013BSP10 \u7684\u80de\u5185 Neu Ac \u542b\u91cf\u5728 230\u2013635 \u03bcg\/m L\u4e4b\u95f4\u53d8\u5316\u3002\u5176\u4e2d\uff0cBSP1\u3001BSP2\u3001BSP7 \u548c BSP9\u4e4b\u95f4\u7684\u80de\u5185 Neu Ac\u542b\u91cf\u5dee\u5f02\u663e\u8457\uff0c\u4e14\u80de\u5185 Neu Ac\u6d53\u5ea6\u8303\u56f4\u66f4\u5e7f(230\u2013635 \u03bcg\/m L)\uff0c\u53ef\u7528\u4e8e\u540e\u7eed\u54cd\u5e94 Neu Ac \u751f\u7269\u4f20\u611f\u5668\u7684\u6027\u80fd\u9a8c\u8bc1\u3002<\/p>\n\n\n\n 2.2 \u54cd\u5e94 Neu Ac \u751f\u7269\u4f20\u611f\u5668\u7684\u8bbe\u8ba1\u4e0e\u6784\u5efa<\/strong><\/p>\n\n\n\n \u57fa\u4e8e\u8f6c\u5f55\u56e0\u5b50\u7684\u751f\u7269\u4f20\u611f\u5668\u901a\u5e38\u5305\u62ec 3 \u4e2a\u90e8\u5206\uff1a\u54cd\u5e94\u7279\u5b9a\u4ee3\u8c22\u7269\u7684\u8f6c\u5f55\u56e0\u5b50\uff0c\u542b\u6709\u8f6c\u5f55\u56e0\u5b50\u7ed3\u5408\u4f4d\u70b9\u7684\u542f\u52a8\u5b50\u4ee5\u53ca\u53d7\u5176\u8c03\u63a7\u8868\u8fbe\u7684\u62a5\u544a\u57fa\u56e0[25-26]\u3002\u54cd\u5e94 Neu Ac \u751f\u7269\u4f20\u611f\u5668\u7684\u539f\u7406\u5982\u56fe 3A \u6240\u793a\uff0c\u5f53\u80de\u5185\u4e0d\u5b58\u5728 Neu Ac \u65f6\uff0cBbr_Nan R\u4f5c\u4e3a\u963b\u904f\u86cb\u767d\u7ed3\u5408\u5728\u5176\u7ed3\u5408\u4f4d\u70b9\u4e0a\uff0c\u6291\u5236\u54cd\u5e94\u542f\u52a8\u5b50\u8c03\u63a7\u7684\u62a5\u544a\u57fa\u56e0\u7684\u8868\u8fbe\uff1b\u5f53\u80de\u5185\u5b58\u5728Neu Ac \u65f6\uff0c\u80de\u5185 Neu Ac \u4e0e Bbr_Nan R \u7684\u7ed3\u5408\u4f1a\u5f71\u54cd Bbr_Nan R \u7684\u963b\u904f\u6d3b\u6027\uff0c\u4ece\u800c\u6fc0\u6d3b\u54cd\u5e94\u542f\u52a8\u5b50\u7684\u8868\u8fbe\u3002<\/p>\n\n\n\n \u56e0\u6b64\uff0c\u9996\u5148\u8bbe\u8ba1\u4e86\u542b\u6709 Nan R \u7ed3\u5408\u4f4d\u70b9\u7684\u6742\u5408\u542f\u52a8\u5b50\u3002\u5982\u56fe 3B \u6240\u793a\uff0c\u5c06 Bbr_Nan R \u7279\u5f02\u6027\u7ed3\u5408\u4f4d\u70b9\u653e\u5728\u4e86 6 \u4e2a\u7ec4\u6210\u578b\u542f\u52a8\u5b50 Pveg\u3001P43\u3001Pm1\u3001P214\u3001P566\u3001P535[27-28]\u7684\u201335 \u533a\u524d\u3001\u201335 \u533a\u548c\u201310 \u533a\u4e2d\u95f4\u3001\u201310 \u533a\u540e\uff0c\u8bbe\u8ba1\u4e86 21 \u4e2a\u5e26\u6709 Bbr_Nan R \u7ed3\u5408\u4f4d\u70b9\u7684\u6742\u5408\u542f\u52a8\u5b50 N1\u2013N21 (\u5e8f\u5217\u5982\u8868 4 \u6240\u793a)\u3002\u4ee5 p HT01 \u8d28\u7c92\u4e3a\u8f7d\u4f53\uff0c\u5206\u522b\u7528\u6742\u5408\u542f\u52a8\u5b50\u8868\u8fbe GFP\uff0c\u5c06\u6784\u5efa\u6210\u529f\u7684\u8d28\u7c92\u8f6c\u5316 BS-nan T \u83cc\u682a\u4ee5\u68c0\u6d4b\u6742\u5408\u542f\u52a8\u5b50\u6d3b\u6027\u3002\u5982\u56fe 4 \u6240\u793a\uff0c\u867d\u7136\u5927\u591a\u6570\u6742\u5408\u542f\u52a8\u5b50\u5728\u63d2\u5165 Nan R\u7684\u7ed3\u5408\u5e8f\u5217\u540e\u6d3b\u6027\u660e\u663e\u964d\u4f4e\u751a\u81f3\u5931\u53bb\u6d3b\u6027\uff0c\u4f46\u662f\u6742\u5408\u542f\u52a8\u5b50 N1\u3001N2\u3001N10\u3001N11\u3001N14\u3001N16\u3001N17 \u663e\u793a\u51fa\u4e86\u63a5\u8fd1\u751a\u81f3\u8d85\u8fc7\u5f3a\u7ec4\u6210\u578b\u542f\u52a8\u5b50P566 \u7684\u6d3b\u6027\uff0c\u56e0\u6b64\uff0c\u9009\u62e9\u8fd9\u4e9b\u6742\u5408\u542f\u52a8\u5b50\u7528\u4e8e\u540e\u7eed\u54cd\u5e94 Neu Ac 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