Construction of shuttle expression plasmid and stable expression of foreign gene in mycobacteria and E. coli.
By employing the pUC19 as a backbone, the regulatory and signal sequences which encode kanamycin resistance, and mycobacterial plasmid origin of replication (oriM) were cloned into the pUC19. The recombinant E. Coli-mycobacteria shuttle expression plasmid pBCG-8000 was constructed. The pBCG-8000 was able to replicate in both E. Coli and mycobacteria (including BCG) systems, and to confer stable kanamycin resistance upon transformants. The study should facilitate the development of BCG and other mycobacteria into multivalent vaccine vectors.
Huangfu
Y M
YM
Department of Medical Molecular Biology, Tongji Medical University, Wuhan.
Zhang
D J
DJ
Cheng
J Z
JZ
Qian
M
M
Liang
J Q
JQ
Li
D
D
D001500:BCG Vaccine / Q000235:genetics; D001483:Base Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics*; D008969:Molecular Sequence Data; D009161:Mycobacterium / Q000235:genetics*; D010957:Plasmids / Q000235:genetics*; D014169:Transformation, Bacterial
D001500:BCG Vaccine