刺糖多孢菌中铵载体蛋白基因<em>amtS</em>的克隆及功能研究

doi:10.13523/j.cb.20150204

中国生物工程杂志

2015, Vol. 35

Issue (2): 25-30 DOI: 10.13523/j.cb.20150204

研究报告

刺糖多孢菌中铵载体蛋白基因amtS的克隆及功能研究

陶文娜, 夏立秋, 丁学知, 唐滢

湖南师范大学生命科学学院微生物分子生物学国家重点实验室培育基地长沙 410081

Cloning and Function Study of amtS Gene from Saccharopolyspora spinosa

TAO Wen-na, XIA Li-qiu, DING Xue-zhi, TANG Ying

College of Life Science, Hunan Normal University, Hunan Provincial Key Laboratory of Microbial Molecular Biology-State Key Laboratory Breeding Base of Microbial Molecular Biology, Changsha 410081, China

全文: PDF(916 KB) HTML

摘要：

目的:铵离子是细胞内合成各种核酸、氨基酸和辅助因子等含氮化合物的重要原料之一。微生物细胞膜上的铵载体蛋白介导了铵离子的转运。通过异源表达刺糖多孢菌中铵载体蛋白基因,研究其对链霉菌产孢能力和次级代谢产物产量的影响。方法:从刺糖多孢菌S04-41菌株中克隆铵载体蛋白基因amtS,通过接合转移导入天蓝色链霉菌M145和变铅青链霉菌TK24中,分析比较amtS基因的异源表达对其产孢能力和次级代谢产物产量的影响。结果:天蓝色链霉菌重组菌株M145/pMF-amtS和变铅青链霉菌重组菌株TK24/pMF-amtS中放线紫红素的产量分别提高了2.85倍和30.02倍。结论:刺糖多孢菌中的铵载体蛋白能够提高链霉菌中次生代谢产物的产量,为进一步研究该基因的功能与对刺糖多孢菌中多杀菌素合成的作用奠定了重要基础。

关键词： 铵转运酶蛋白; 链霉菌; 异源表达; 接合转移

Abstract:

Objective: Ammonium ion is one of the precursors for biosynthesis of nucleic acids, amino acids and cofactors. The transportation of NH₄⁺ through the cell plasma membrane is mediated by ammonium carrier protein. The ammonium carrier protein encoding gene of Saccharopolyspora spinosa was cloned, and its effects on the production of secondary metabolites of Streptomyces coelicolor and Streptomyces lividans were evaluated. Methods:The amtS gene of S. spinosa was cloned into E. coli-Streptomyces shuttle vector pMF and transferred into S. coelicolor M145 and S. lividans TK24 by conjugation. The phenotypes of the recombinants were compared with the parental strains, and the effects of amtS overexpression on the production of secondary metabolites were analyzed. Results:The yields of actinorhodin and undecylprodigiosin in both M145/pMF-amtS and TK24/pMF-amtS were greatly improved. The yields of actinorhodin were elevated by 2.85 and 30.02 times, respectively. Conclusion:Heterologous expression of amtS from Saccharopolyspora spinosa could improve the production of secondary metabolites, which provided the important basis for its function study in Saccharopolyspora spinosa.

Key words: Ammonium carrier protein Streptomyces Heterologous expression Conjugation

收稿日期: 2014-12-03 出版日期: 2015-02-25

ZTFLH:

Q819

基金资助:

国家"973"计划专项(2012CB722301)、国家"863"计划(2011AA10A203)、湖南省自然科学基金(2015JJ6071)、湖南省协同创新中心项目(20134486)资助项目

通讯作者: 夏立秋 E-mail: xialq@hunnu.edu.cn

	服务
	把本文推荐给朋友
	加入引用管理器
	E-mail Alert
	RSS
	作者相关文章

引用本文:

陶文娜, 夏立秋, 丁学知, 唐滢. 刺糖多孢菌中铵载体蛋白基因amtS的克隆及功能研究[J]. 中国生物工程杂志, 2015, 35(2): 25-30.

TAO Wen-na, XIA Li-qiu, DING Xue-zhi, TANG Ying. Cloning and Function Study of amtS Gene from Saccharopolyspora spinosa. China Biotechnology, 2015, 35(2): 25-30.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20150204 或 https://manu60.magtech.com.cn/biotech/CN/Y2015/V35/I2/25

[1] Tang Y, Xia L, Ding X, et al. Duplication of partial spinosyn biosynthetic gene cluster in Saccharopolyspora spinosa enhances spinosyn production. FEMS Microbiol Lett, 2011,325(1):22-29.

[2] Waldron C,Matsushima P,Rosteck P R. Cloning and analysis of the spinosad biosynthetic gene cluster of Saccharopolyspora spinosa. Chemistry and Biology,2001,8(5):487-499.

[3] Luo Y, Ding X, Xia L, et al.Comparative proteomic analysis of Saccharopolyspora spinosa SP06081 and PR2 strains reveals the differentially expressed proteins correlated with the increase of spinosad yield. Proteome Sci,2011, 9:40.

[4] Luo Y, Kou X, Ding X, et al.Promotion of spinosad biosynthesis by chromosomal integration of the Vitreoscilla hemoglobin gene in Saccharopolyspora spinosa. Sci China Life Sci, 2012,55(2):172-180.

[5] 邓若磊, 徐海荣, 曹云飞, 等.植物吸收铵态氮的分子生物学基础. 植物营养与肥料学报,2007,13 (3):512-519. Deng R L, Xu H R, Cao Y F, et al. The molecular basis of ammonium transporters in plant. Plant Nutrition and Fertilizer Science, 2007,13 (3):512-519.

[6] Van Wezel G P, Van der Meulen J, Kawamoto S, et al. SsgA is essential for sporulation of Streptomyces coelicolor A3(2) and affects hyphal development by stimulating septum formation. J Bacteriol, 2000,182(20):5653-5662.

[7] Shima J, Hesketh A, Okamoto S, et al. Induction of actinorhodin production by rpsL (encoding ribosomal protein S12) mutations that confer streptomycin resistance in Streptomyces lividans and Streptomyces coelicolor A3(2). J Bacteriol,1996,178(24):7276-7284.

[8] Kesier T, Bibb M J, Butter M J, et al. Practical Streptomyces Genetics. Norwich (United K):John Innes Foundation,2000, 488(1):25-37.

[9] Chinping Chng, Amy M L, Jonathan A Vroom, et al. A key developmental regulator controls the synthesis of the antibiotic erythromycin in Saccharopolyspora erythraea. PNAS,2008, 105,(32):11346-11351.

[10] Zhang Q, Zhu B Q, Hu H F. Activated antibiotic production by inducing resistance to capreomycin in Streptomyces lividans and Streptomyces coelicolor.Chinese Journal of Natural Medicines,2008,6:57-62.

[1]	冯宝琪,冯娇,张苗,刘洋,曹睿,尹涵之,齐凤仙,李子龙,尹守亮. 利用Tn5型转座突变系统筛选高产阿维菌素菌株^*[J]. 中国生物工程杂志, 2021, 41(7): 32-41.
[2]	王珊,薛正莲,孙俊峰,王芳,周健,刘艳,王洲. 盐增强培养对弗氏链霉菌产新霉素的影响[J]. 中国生物工程杂志, 2021, 41(7): 22-31.
[3]	李开秀,司维. 间充质干细胞来源的外泌体治疗炎症性肠病研究进展^*[J]. 中国生物工程杂志, 2021, 41(7): 66-73.
[4]	王优蓓,郭思妤,常碧博,叶蕊芳,花强. 螺旋链霉菌遗传操作系统-接合转移体系的建立[J]. 中国生物工程杂志, 2021, 41(2/3): 45-52.
[5]	饶海密,梁冬梅,李伟国,乔建军,财音青格乐. 真菌芳香聚酮化合物的合成生物学研究进展^*[J]. 中国生物工程杂志, 2020, 40(9): 52-61.
[6]	位薇,常保根,王英,路福平,刘夫锋. Tau蛋白核心片段306~378的异源表达、纯化及聚集特性验证^*[J]. 中国生物工程杂志, 2020, 40(5): 22-29.
[7]	李吉萍,包昌杰,陈光,张斯童. 木聚糖酶异源表达的研究进展 ^*[J]. 中国生物工程杂志, 2019, 39(7): 91-99.
[8]	吴果果,宋淑婷,岳荣,张晶,关莹,王玥,刘宝爱,吕学敏,魏建军,张会图. 反向筛选标记基因upp在杀真菌链霉菌遗传改造中的应用 ^*[J]. 中国生物工程杂志, 2019, 39(11): 78-86.
[9]	史超硕,李登科,曹雪,袁航,张钰文,于江悦,路福平,李玉. 两个不同启动子及其组合对碱性蛋白酶AprE异源表达的影响 ^*[J]. 中国生物工程杂志, 2019, 39(10): 17-23.
[10]	陈子晗,周海胜,尹新坚,吴坚平,杨立荣. **Amphibacillus xylanus谷氨酸脱氢酶基因工程菌培养条件优化 ^***[J]. 中国生物工程杂志, 2019, 39(10): 58-66.
[11]	李博, 梁楠, 刘夺, 刘宏, 王颖, 肖文海, 姚明东, 元英进. 合成8二甲基异戊烯基柚皮素的人工酿酒酵母菌株构建[J]. 中国生物工程杂志, 2017, 37(9): 71-81.
[12]	宋佳雯, 田苏, 张玉如, 王志珍, 常忠义, 高红亮, 步国建, 金明飞. 基因组重排筛选高产谷氨酰胺转胺酶菌株[J]. 中国生物工程杂志, 2017, 37(9): 105-111.
[13]	李丹, 黄鹤. 纳米抗体异源表达的研究进展[J]. 中国生物工程杂志, 2017, 37(8): 84-95.
[14]	田淑翠, 牛延宁, 常忠义, 高红亮, 步国健, 金明飞. 常压室温等离子体(ARTP)诱变茂源链霉菌菌株[J]. 中国生物工程杂志, 2016, 36(9): 47-53.
[15]	尹守亮, 林智炜, 张玉秀, 王为善, 史明欣, 杨克迁. 工程改造龟裂链霉菌提高土霉素产量[J]. 中国生物工程杂志, 2016, 36(7): 72-82.

Viewed

Full text

Abstract

Cited

Shared

Discussed