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KMID : 0880220060440060671
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Journal of Microbiology
2006 Volume.44 No. 6 p.671 ~ p.673
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Construction of an Escherichia-Pseudomonas Shuttle Vector Containing an Aminoglycoside Phosphotransferase Gene and a lacZ’’’’ Gene for α-Complementation
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Lee Bheong-Uk
Hong Ja-Heon
Kahng Hyung-Yeel
Oh Kye-Heon
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Abstract
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A new 4.87 kb Escherichia-Pseudomonas shuttle vector has been constructed by inserting a 1.27 kb DNA fragment with a replication origin of a Pseudomonas plasmid pRO1614 into the 3.6 kb E. coli plasmid pBGS18. This vector, designated pJH1, contains an aminoglycoside phosphotransferase gene (aph) from Tn903, a lacZ’’’’ gene for α-complementation and a versatile multiple cloning site possessing unique restriction sites for EcoRI, SacI, KpnI, SmaI, BamHI, XbaI, SalI, BspMI, PstI, SphI, and HindIII. When pJH1 was transformed into E. coli DH5α and into P. putida HK-6, it was episomally and stably maintained in both strains. In addition, the enhanced green fluorescent protein (EGFP) gene which was transcriptionally cloned into pJH1 rendered E. coli cells fluorescence when its transformants were illuminated at 488 nm.
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KEYWORD
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Pseudomonas, shuttle vector, kanamycin-resistant gene, cloning
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