固氮链霉菌Streptomyces chartreusi WZS021接合转移系统的建立及优化

【目的】建立和优化固氮链霉菌Streptomyces chartreusi WZS021（简称菌株WZS021,下同）的接合转移体系,以丰富固氮链霉菌基因片段转移技术。【方法】以大肠杆菌Escherichia coli ET12567（pUZ8002）为供体菌,携带整合型质粒pSET152的菌株WZS021为受体菌,进行菌株WZS021接合转移。【结果】选择高氏一号培养基为菌株WZS021接合转移培养基,50℃热激10 min,供受比为1∶10,涂布20.0 mg/L安普霉素、接合转移12 h后覆盖抗生素,添加MgCl2终浓度为5.0 mmol/L时接合转移效率最高,达3.24×10-6。【结...

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Bibliographic Details
Published in南方农业学报 Vol. 48; no. 4; pp. 581 - 586
Main Author 王震 庞妃 顾彩彩 王露蓉 邢永秀 杨丽涛 李杨瑞
Format Journal Article
LanguageChinese
Published 广西大学 农学院/亚热带农业生物资源保护与利用国家重点实验室,南宁,530004%广西大学 农学院/亚热带农业生物资源保护与利用国家重点实验室, 南宁 530004 2017
广西农业科学院 甘蔗研究所/中国农业科学院 甘蔗研究中心/农业部广西甘蔗生物技术与遗传改良重点实验室/广西甘蔗遗传改良重点实验室, 南宁 530007
Subjects
pSET152
优化
固氮链霉菌WZS021
接合转移
固氮链霉菌WZS021
pSET152
optimization
优化
接合转移
transconjugation
Streptomyces chartreusi WZS021
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ISSN2095-1191
DOI10.3969/j.issn.2095-1191.2017.04.003

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Summary:【目的】建立和优化固氮链霉菌Streptomyces chartreusi WZS021（简称菌株WZS021,下同）的接合转移体系,以丰富固氮链霉菌基因片段转移技术。【方法】以大肠杆菌Escherichia coli ET12567（pUZ8002）为供体菌,携带整合型质粒pSET152的菌株WZS021为受体菌,进行菌株WZS021接合转移。【结果】选择高氏一号培养基为菌株WZS021接合转移培养基,50℃热激10 min,供受比为1∶10,涂布20.0 mg/L安普霉素、接合转移12 h后覆盖抗生素,添加MgCl2终浓度为5.0 mmol/L时接合转移效率最高,达3.24×10-6。【结论】通过确定适用于菌株WZS021接合转移条件建立的菌株WZS021遗传转化体系,有助于今后插入标记基因确定该菌的固氮定殖位点及导入外源基因的操作。
Bibliography:45-1381/S
Streptomyces ehartreusi WZS021 ; transconjugation ; optimization ; pSET152
WANG Zhen1, PANG Fei1, GU Cai-cai1, WANG Lu-rong1, XING Yong-xiu1, YANG Li-tao1,2, LI Yang-rui1,2（1.College of Agriculture, Guangxi University / State Key Laboratory of Conservation and Utilization of Subtropical Agrobioresources, Nanning 530004, China; 2Sugarcane Research Institute,Guangxi Academy of Agricultural Sciences/Sugarcane Research Center, Chinese Academy of Agricultural Sciences/Guangxi Key Laboratory of Sugarcane Biotechnology and Genetic Improvement, Ministry of Agriculture/Guangxi Key Laboratory of Sugarcane Genetic hnprovement, Nanning 530007, China）
[Objective ]The present study was conducted to establish and optimize transconjugation system of Streptomyces chartrezsis WZS021 （hereinafter referred to as strain WZS021 ）,in order to enrich strain WZS021 gene fragment transfer technology. [ Method ] As donor strain, Escberichia coli ET12567 （pUZ8002） contained integrative plasmid pSET152. Strain WZS021 was recipient
ISSN:2095-1191
DOI:10.3969/j.issn.2095-1191.2017.04.003