| 卡介苗菌株和结核分枝杆菌H37Ra菌株原生质体的制备研究 |
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| 引用本文： | 樊超,;姜晓霞,;张锋,;王霞,;吴芳,;章乐,;吴江东,;张春军,;张辉,;梁晨,;庄睿,;李文娟,;张万江. 卡介苗菌株和结核分枝杆菌H37Ra菌株原生质体的制备研究[J]. 石河子大学学报(自然科学版), 2014, 0(2): 175-179 |
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| 作者姓名： | 樊超, 姜晓霞, 张锋, 王霞, 吴芳, 章乐, 吴江东, 张春军, 张辉, 梁晨, 庄睿, 李文娟, 张万江 |
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| 作者单位： | [1]石河子大学医学院/新疆地方与民族高发病教育部重点实验室,石河子832002; [2]石河子大学医学院第一附属医院功能科,石河子832008; [3]新疆医科大学第五附属医院烧伤整形科,乌鲁木齐830011; [4]阿克苏地区第二人民医院检验科,阿克苏843000 |
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| 基金项目： | 国家自然科学基金项目（81260261,81160192,81260241,81160001）,新疆兵团医药卫生专项项目（2012BA022） |
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| 摘 要： | 分别以卡介苗菌株(BCG)及结核分枝杆菌国际标准无毒株H37Ra菌株(H37Ra)为受体菌,制备上述两种菌株的原生质体,同时通过优化细菌菌龄、酶解浓度、酶解温度以及酶解时间等影响因素,探索出制备原生质体形成及再生的最优条件。结果显示:摸索出制备BCG和H37Ra菌株的原生质体条件:在对数生长期的两亲本菌株,经0.01 mol/L EDTA,0.01%β-巯基乙醇溶液预处理;酶解浓度为12 mg/mL,酶解温度为37℃,酶解时间为5 h,可制备出活性较高的两种菌株的原生质体。由此可知,成功制备了BCG与H37Ra菌株的原生质体,并能在高渗固体培养基上再生。本实验为进一步研究该两种菌株原生质体融合试验奠定了基础。
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| 关 键 词： | 原生质体制备 BCG菌株 结核分枝杆菌H37Ra菌株 影响因素 |
Study on Formation of Protoplast of BCG Strains and the Mycobacterium Tuberculosis H37Ra Strains |
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| Affiliation: | FAN Chao, JIANG Xiaoxia, ZHANG Feng, WANG Xia, WU Fang, ZHANG Le, WU Jiangdong, ZHANG Chunjun, ZHANG Hui, LING Chen, ZHUANG Rui, LI Wenjuan, ZHANG Wanjiang (1 Key Laboratory of Xinjiang Endemic and Ethnic Diseases/Department of Immunology,School of Medicine Shihezi University,Shihezi 832002,China; 2 Department of Function,the First Affiliated Hospital,School of Medicine,Shihezi University,Shihezi 832002,China; 3. Burns and Plastic Surgery,the Fifth Affiliated Hospital of Xinjiang Medical University,Urumchi 850011,China; 4 Clinical Laboratory,the Second People＇s Hospital of Xinjiang Aksu Region,Aksu 843000,China) |
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| Abstract: | To explore the formation of protoplasts from BC G strains （hereinafter referred to as the BCG） and the Mycobacterium tuberculosis H37Ra strains （hereinafter referred to as the H37Ra strain）.Optimal conditions for the formation and regeneration of protoplasts were studied via optimizing of the fungus age,enzyme solution concentration, temperature and action time,the optimal conditions of the formation of BCG and H37Ra strains protoplasts were proved to be as follows：the logarithmic phase of two parent strain, pretreated with 0.01mol/L EDTA and 0.01% β-mercaptoethanol solution,under enzyme concentration 12mg/mL,enzymolysis temperature 37 ℃ and enzymatic hydrolysis time 5h,that can achieve high activity protoplasts.The two protoplasts,successfully obtained from BCG strain and Mycobacterium tuberculosis H37Ra strain,possess the ability of regeneration in hypertonic solid medium,which laid the foundation for further protoplasts fusion experiment. |
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| Keywords: | Formation of Protoplast BCG strains Mycobacterium tuberculosis H37Ra strains Formation conditions |
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