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中国生物工程杂志

China Biotechnology
China Biotechnology  2015, Vol. 35 Issue (5): 81-86    DOI: 10.13523/j.cb.20150512
    
Construction of an Inducible Expression Shuttle Vector for Ketogulonigenium vulgare Allowing Tightly Regulable Gene Expression
LUO Wan-yue, LI Tian-ming, YU Ying, XU Mei-xue, YI Hong
Hebei University of Science and Technology, Shijiazhuang 050018, China
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Abstract  

The genes encoding tet repressor proteins and Ketogulonigenium vulgare sorbose dehydrogenase operator which inserted operon teto were synthesized by overlapping PCR. The shuttle vector of tetracycline induced expression with broad host plasmid pBBR1MCS-5 was constructed and transformed Ketogulonigenium vulgare. The resistance of kanamycin was regulatively expressed after getting the positive recombinant strains. The results show that the cultivation of recombinant strains could grow in the medium containing kanamycin where added 0.4mg/ml tetracycline inducers after 2 hours, while they can't grow in the medium containing kanamycin where didn't add tetracycline inducers. The optimal induction of tetracycline concentration determined was 0.6mg/ml.



Key wordsKetogulonigenium vulgare      Gene synthesis      Shuttle vector      Tetracycline inducible     
Received: 16 January 2015      Published: 25 May 2015
ZTFLH:  Q789  
Cite this article:

LUO Wan-yue, LI Tian-ming, YU Ying, XU Mei-xue, YI Hong. Construction of an Inducible Expression Shuttle Vector for Ketogulonigenium vulgare Allowing Tightly Regulable Gene Expression. China Biotechnology, 2015, 35(5): 81-86.

URL:

https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20150512     OR     https://manu60.magtech.com.cn/biotech/Y2015/V35/I5/81


[1] Yang F, Jia Q, Xiong Z, et al. Complete genome analysis of Ketogulonigenium sp. WB0104. Chinese Science Bulletin, 2006, 51(8): 941-945

[2] Xiong X, Han S, Wang J, et al. Complete genome sequence of the bacterium Ketogulonicigenium vulgare Y25. Journal of Bacteriology, 2011, 193(1): 315-316.

[3] Liu W, Zhang L, Yang J, et al. Reconstruction and analysis of a genome-scale metabolic model of the vitamin C producing industrial strain Ketogulonicigenium vulgare WSH-001.Journal of Biotechnology, 2012,161 (1):42-48.

[4] Berens C, Hillen W. Gene regulation by tetracyclines. European Journal of Biochemistry, 2003, 270(15): 3109-3121.

[5] Geissendrfer M, Hillen W. Regulated expression of heterologous genes in Bacillus subtilis using the Tn10 encodedtet regulatory elements. Applied Microbiology and Biotechnology, 1990, 33(6): 657-663.

[6] Ji Y, Zhang B, Van S F, et al. Identification of critical staphylococcal genes using conditional phenotypes generated by antisense RNA. Science, 2001, 293(5538): 2266-2269.

[7] Lutz R, Bujard H. Independent and tight regulation of transcriptional units in Escherichia coli via the LacR/O, the TetR/O and AraC/I1-I2 regulatory elements. Nucleic Acids Research, 1997, 25(6): 1203-1210.

[8] Qian F, Pan W. Construction of a tetR-integrated Salmonella enterica serovar Typhi CVD908 strain that tightly controls expression of the major merozoite surface protein of Plasmodium falciparum for applications in human vaccine production. Infection and Immunity, 2002, 70(4): 2029-2038.

[9] 李建,宁静,陈西锐,等. 小鼠H1foo基因四环素诱导表达系统的构建与表达. 畜牧兽医学报,2014,45(5):757-762. Li J, Ning J, Chen X R, et al. Construction of tetracycline-inducible expression system of H1foo gene in mouse.Acta Veterinaria et Zootechnica Sinica, 2014,45(5):757-762.

[10] 解宇,汪亮,张冬杰,等.四环素诱导的猪黑素皮质激素受体-4基因真核表达载体的构建.中国畜牧兽医,2013,40(1):13-16. Xie Y, Wang L, Zhang D J, et al. Construction of tetracycline-inducible MC4R gene eukaryotic expression vector in pig. Chinese Animal Husbandry and Veterinary Medicine, 2013,40(1):13-16.

[11] 吴亮,薛兰兰,王晓,等.用四环素诱导调控表达系统构建弓形虫脂肪酸合成酶FABZ缺陷株.中国寄生虫学与寄生虫病杂志,2014,32(4):264-267. Wu L, Xue L L, Wang X, et al. Construction of β-hydroxyacyl-acyl carrier protein dehydratase mutant of Toxoplasma gondii by tetracycline inducible expression system,Chinese Journal of Parasitology and Parasitic Diseases, 2014,32(4):264-267.

[12] Sabine E, Guo X V, Christopher M H. Controlling gene expression in mycobacteria with anhydrotetracycline and Tet repressor. Nucleic Acids Research,2005,3(33):e21

[13] Miyazaki T, Sugisawa T, Hoshino T. Pyrroloquinoline quinone-dependent dehydrogenases from Ketogulonicigenium vulgare catalyze the direct conversion of L-sorbosone to L-ascorbic acid. Applied and Environmental Microbiology, 2006, 72(2): 1487-1495.

[14] Knauf V C, Nester E W. Wide host range cloning vectors: a cosmid clone bank of an Agrobacterium Ti plasmid. Plasmid, 1982, 8(1): 45-54.

[15] Kovach M E, Elzer PH, Hill D S, et al. Four new derivatives of the broad-host-range cloning vector pBBR1MCS, carrying different antibiotic-resistance cassettes. Gene, 1995, 166(1): 175-176.

[16] Fu S, Zhang W, Guo A, et al. Identification of promoters of two dehydrogenase genes in Ketogulonicigenium vulgare DSM 4025 and their strength comparison in K. vulgare and Escherichia coli. Applied Microbiology and Biotechnology, 2007, 75(5): 1127-1132..

[17] Tahlan K, Ahn S K, Sing A, et al. Initiation of actinorhodin export in Streptomyces coelicolor. Molecular Microbiology, 2007, 63(4): 951-961.

[18] Wright K J, Seed PC, Hultgren S J. Development of intracellular bacterial communities of uropathogenic Escherichia coli depends on type 1 pili. Cellular Microbiology, 2007, 9(9): 2230-2241.

[19] Guo X V, Monteleone M, Klotzsche M, et al. Silencing essential protein secretion in Mycobacterium smegmatis by using tetracycline repressors. Journal of Bacteriology, 2007, 189(13): 4614-4623.

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