| 新型抗结核分枝杆菌药物筛选模型的建立 |
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| 引用本文： | 张磊磊,由鹏飞,姚广新,侯爵,张怡轩.新型抗结核分枝杆菌药物筛选模型的建立[J].微生物学杂志,2011,31(1):57-62. |
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| 作者姓名： | 张磊磊 由鹏飞 姚广新 侯爵 张怡轩 |
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| 作者单位： | 1. 沈阳药科大学,生物制药教研室,辽宁,沈阳,110016
2. 沈阳药科大学,生物制药教研室,辽宁,沈阳,110016;沈阳药科大学,生物学教学实验中心,辽宁,沈阳,110016 |
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| 摘 要： | 为建立基于酶水平和细胞水平的新型抗结核分枝杆菌（Mycobacterium tuberculosis）药物的筛选模型,以M.tuberculosis H37Rv基因组DNA为模板,PCR特异性扩增异柠檬酸裂解酶（ICL）基因,构建表达载体,在E.coli BL21（DE3）中高效表达,使用N i2＋亲和层析柱纯化重组ICL,检测其活性。优化ICL酶反应条件,考察待筛选样品溶剂对酶活性的影响,建立ICL抑制剂酶水平筛选模型;考察与优化耻垢分枝杆菌（Mycobacterium smegma）在以乙酸盐为唯一碳源的培养基中的生长状况,建立基于M.sm egma的乙醛酸代谢途径抑制剂的细胞水平筛选模型;利用上述2种筛选模型对1 060种可能具有拮抗活性的微生物代谢样品进行初筛和复筛,两者筛选结果正相关性较好。
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| 关 键 词： | 结核分枝杆菌 异柠檬酸裂解酶 耻垢分枝杆菌 乙醛酸途径 筛选模型 |
Establishment of New Screening Models for Anti-Mycobacterium tuberculosis Medicines |
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| ZHANG Lei-lei,YOU Peng-fei,YAO Guang-xin,HOU Jue,ZHANG Yi-xuan.Establishment of New Screening Models for Anti-Mycobacterium tuberculosis Medicines[J].Journal of Microbiology,2011,31(1):57-62. |
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| Authors: | ZHANG Lei-lei YOU Peng-fei YAO Guang-xin HOU Jue ZHANG Yi-xuan |
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| Institution: | ZHANG Lei-lei1,YOU Peng-fei1,2,YAO Guang-xin1,HOU Jue1,ZHANG Yi-xuan1,2(1.Teach.& Res.Div.of Bio-Pharm.,2.Biol.& Exp Teach.Ctr.,Shenyang Pharm.Uni.,Shenyang 110016) |
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| Abstract: | Mycobacterium tuberculosis strain H37Rv genomic DNA was adopted as a template to establish a screening model targeting to isocitrate lyase(ICL) and glyoxylate pathway for anti-TB drugs at enzymatic and cellular levels.ICL gene was amplified by polymerase chain reaction(PCR) and constructed an expression vector and highly expressed in E.coli BL21(DE3).The recombinant ICL was purified with Ni2+-NTA resin affinity chromatography and detected for its activity.The ICL enzymatic reaction conditions were optimized... |
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| Keywords: | Mycobacterium tuberculosis isocitrate lyase Mycobacterium smegmatis glyoxylate pathway screening model |
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