[Construction of Escherichia coli-Mycobacteria shuttle plasmid and the stable expression of human interleukin-2 in BCG and Escherichia coli].
OBJECTIVE
To construct and identify Escherchia coli (E. coli)-Mycobacteria shuttle plasmid and to detect stable expression of foreign gene in E. coli and BCG.


METHODS
With a genetic engineering technique to construct the E. coli-Mycobacteria shuttle plasmid, the human interleukin-2 (IL-2) gene was electrophoreted into BCG with recombinant plasmid PZSIII-I and positive clones selected using polymerase chain reaction (PCR) technique. The expression of foreign gene of human IL-2 in BCG was identified by ELISA and SDS-PAGE.


RESULTS
Human IL-2 cytokine was steadily expressed in recombinant BCG and E. coli and could secrete outside the cell.


CONCLUSIONS
M. bovis BCG recombinant constructed can produce and secrete the human IL-2. A secretion of the active cytokine was accomplished through the combined use of the BCG HSP65 promoter and a secretion signal from the BCG Ag-85B. The BCG HSP65 promoter is active in both BCG as well as E. coli which can not secrete foreign protein.
Zeng
X
X
Central Laboratory, Second Affiliated Hospital, Guangzhou Chinese Traditional Medicine University.
Yang
M
M
Qi
X
X
D001500:BCG Vaccine / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular / Q000379:methods; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D005822:Genetic Vectors; D006801:Humans; D007376:Interleukin-2 / Q000096:biosynthesis* / Q000235:genetics; D009161:Mycobacterium / Q000235:genetics*; D010957:Plasmids / Q000235:genetics*
D001500:BCG Vaccine; D007376:Interleukin-2