Construction of an Escherichia-Pseudomonas shuttle vector containing an aminoglycoside phosphotransferase gene and a lacZ'' Gene for alpha-complementation.
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 alpha-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 DH5alpha 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.
Lee
Bheong-Uk
BU
Division of Biological Sciences, Kosin University, Busan 606-701, Republic of Korea.
Hong
Ja-Heon
JH
Kahng
Hyung-Yeel
HY
Oh
Kye-Heon
KH
D000900:Anti-Bacterial Agents / Q000494:pharmacology; D003001:Cloning, Molecular; D024881:Drug Resistance, Bacterial; D004925:Escherichia / Q000201:enzymology / Q000235:genetics*; D005816:Genetic Complementation Test; D005822:Genetic Vectors / Q000235:genetics*; D007612:Kanamycin / Q000494:pharmacology; D019868:Kanamycin Kinase / Q000235:genetics*; D007763:Lac Operon; D010957:Plasmids; D011549:Pseudomonas / Q000201:enzymology / Q000235:genetics*
D000900:Anti-Bacterial Agents; D007612:Kanamycin; D019868:Kanamycin Kinase