| 双歧杆菌原生质体的制备及其转化系统的建立 |
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| 引用本文： | 万翠香,章昭琳,王报贵,魏华.双歧杆菌原生质体的制备及其转化系统的建立[J].食品与发酵工业,2012,38(6):61-65. |
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| 作者姓名： | 万翠香 章昭琳 王报贵 魏华 |
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| 作者单位： | 1. 南昌大学食品科学与技术国家重点实验室,江西南昌330047 南昌大学中德联合研究院,江西南昌330047
2. 南昌大学食品科学与技术国家重点实验室,江西南昌,330047 |
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| 基金项目： | 国家自然科学基金项目“双歧杆菌丝氨酸蛋白酶抑制剂的功能研究”，国家“863”计划专题“具有高黏附、拮抗和抗炎症能力的双歧杆菌的高效筛选和功能基因的定位研究”，国家自然科学基金项目“磁珠法和量子点标记筛选及验证双歧杆菌的粘附蛋白”，2009年江西省主要学科学术和技术带头人培养计划“双歧杆菌Serpin黏附和免疫作用的研究”，江西省教育厅科研基金“新型双歧杆菌功能蛋白Serpin的研究” |
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| 摘 要： | 考察了长双歧杆菌WBL001菌体生长状态、酶解浓度、时长、温度以及不同渗透压稳定剂等因素,获得长双歧杆菌WBL001原生质体制备及再生的优化条件:长双歧杆菌以3%接种量活化至MRS培养基,培养至对数中期(OD600∶1),5μg/mL变溶菌素,37℃孵育30 min,以原生质体稳定液SMM为反应缓冲液,原生质体生成率达到81%,其再生率达到48%;在此基础上,通过聚乙二醇PEG融合,将穿梭表达载体pBAX转入双歧杆菌原生质体,成功建立双歧杆菌转化体系。
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| 关 键 词： | 双歧杆菌 原生质体 再生 变溶菌素 转化 |
Prepareation and Transformation of Bifidobacteria Protoplast |
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| Wan Cui-xiang,Zhang Zhao-lin,Wang Bao-gui,Wei Hua.Prepareation and Transformation of Bifidobacteria Protoplast[J].Food and Fermentation Industries,2012,38(6):61-65. |
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| Authors: | Wan Cui-xiang Zhang Zhao-lin Wang Bao-gui Wei Hua |
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| Affiliation: | 1,2,Gan Yan-yun3 1(State Key Laboratory of Food Science and Technology,Nanchang University,Nanchang 330047,China) 2(Jiangxi-OAI Joint Research Institute,Nanchang 330047,China) |
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| Abstract: | Six factors involved in the preparation and regeneration of Bifidobacteria longum protoplast were investigated,including state of growing cells,Mutanolysin concentration,enzymolysis time,hydrolysis temperature,and osmotic stabilizers.The optimum conditions were as follows: a log phase(19 h) culture of the recipient strain was grown in MRS medium,Mutanolysin concentration was 5 μg/mL,enzymolysis time was 30 min,SMM was used as reaction buffer.Using these optimum conditions,the protoplast formation rate reached 81%,and the ratio of regeneration was 48% in the optimum regeneration medium.On this basis,a Bifidobacteria transformation system of pBAX with PEG was established. |
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| Keywords: | Bifidobacteria protoplasts regeneration Mutanolysin transformation |
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