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| 一个热诱导穿梭表达载体的构建及其在链霉菌中的应用 | |
| 引用本文： | 陶美凤 周秀芬 Tobias Kieser 邓子新.一个热诱导穿梭表达载体的构建及其在链霉菌中的应用[J].生物工程学报,2002,18(4):420-423. |
| 作者姓名： | 陶美凤 周秀芬 Tobias Kieser 邓子新 |
| 作者单位： | 1. 华中农业大学农业部农业微生物重点实验室,武汉,430070 2. 华中农业大学农业部农业微生物重点实验室,武汉,430070;上海交通大学Bio-X生命科学研究中心,上海,200030 3. John Irmes Centre,Norwich,U K |
| 基金项目： | 国家科委863青年科学基金，国家自然科学基金 (No .39970 479,3982 5 10 1)，英国皇家学会资助~~ |
| 摘 要： | 为探索大肠杆菌λ噬菌体表达调控元件在链霉菌中的应用，构建了一个链霉菌大肠杆菌穿梭表达载体pHZ1080，并将来自链霉菌FR-008的聚酮合酶(PKS)基因置于其中的λ噬菌体启动子PR下游，得到表达PKS的穿梭质粒pHZ1067。与在大肠杆菌中一样，该质粒在变铅青链霉菌中也受热诱导表达100kD的PKS蛋白；表达的PKS蛋白可由SDSPAGE和Western-blot实验检测到。PKS在链霉菌中的热诱导表达表明，构建的载体也能用于链霉菌诱导表达外源基因。
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| 关 键 词： | 链霉菌大肠杆菌穿梭表达载体， 热诱导， λ噬菌体启动子， PKS |
| 文章编号： | 1000-3061(2002)04-0420-04 |
| 修稿时间： | 2002年1月23日 |
Construction of a Temperature Inducible Shuttle Expression Vector and Its Application in Streptomyces |
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| Tobias Kieser.Construction of a Temperature Inducible Shuttle Expression Vector and Its Application in Streptomyces[J].Chinese Journal of Biotechnology,2002,18(4):420-423. | |
| Authors: | Tobias Kieser |
| Institution: | Key Laboratory of Agricultural Microbiology, Ministry of Agriculture, Huazhong Agricultural University, Wuhan 430070, China. |
| Abstract: | pHZ1080, an E. coli-Streptomyces shuttle expression vector was constructed in order to explore the utilization of lambda phage regulated expression elements in Streptomyces. A 2.7 kb polyketide synthase (PKS) gene from Streptomyces sp. FR-008 was inserted into downstream of lambda phage promoter (PR) to give the shuttle plasmid, pHZ1067. The PKS protein was expressed in Streptomyces lividans carrying pHZ1067 in a heat-dependent manner, as it did in E. coli. The PKS protein expressed in both hosts with same molecular weight was detected by SDS-PAGE and Western-blot. The successful heat-induced expression of PKS suggested that pHZ1080 was useful and convenient for heat-induced expression of heterologous genes in both E. coli and Streptomyces. |
| Keywords: | PKS |
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