TY - JOUR
T1 - Plasmid vectors designed for high-efficiency expression controlled by the portable recA promoteroperator of Escherichia coli
AU - Masahiro, Shirakawa
AU - Toshiki, Tsurimoto
AU - Kenichi, Matsubara
N1 - Funding Information:
was supported in part by a grant from the Ministry of Education, Science and Culture of Japan.
PY - 1984/4
Y1 - 1984/4
N2 - A novel expression vector using the 236-bp promoter-operator fragment of the recA gene (recApo) of Escherichia coli has been constructed. This DNA fragment contains complete signals for the initiation of RNA synthesis, as well as for regulation by the lexA product, but lacks the coding sequence for the RecA protein. The strength of the recA promoter was examined by assaying ß-galactosidase activity expressed from a cro-lacZ fused gene placed downstream of the promoter. Under noninducing conditions, the promoter was regulated by the LexA protein, and the fused gene was expressed only weakly. Upon induction by nalidixic acid in a recA+ strain, high expression was observed for an extended period. After 5 h under inducing conditions, as much as 11 % of the total cellular protein was cro-lacZ product. The expression level was higher than that from promoters of lac, trp, and λ early genes.
AB - A novel expression vector using the 236-bp promoter-operator fragment of the recA gene (recApo) of Escherichia coli has been constructed. This DNA fragment contains complete signals for the initiation of RNA synthesis, as well as for regulation by the lexA product, but lacks the coding sequence for the RecA protein. The strength of the recA promoter was examined by assaying ß-galactosidase activity expressed from a cro-lacZ fused gene placed downstream of the promoter. Under noninducing conditions, the promoter was regulated by the LexA protein, and the fused gene was expressed only weakly. Upon induction by nalidixic acid in a recA+ strain, high expression was observed for an extended period. After 5 h under inducing conditions, as much as 11 % of the total cellular protein was cro-lacZ product. The expression level was higher than that from promoters of lac, trp, and λ early genes.
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U2 - 10.1016/0378-1119(84)90096-9
DO - 10.1016/0378-1119(84)90096-9
M3 - Article
C2 - 6329913
AN - SCOPUS:0021238591
SN - 0378-1119
VL - 28
SP - 127
EP - 132
JO - Gene
JF - Gene
IS - 1
ER -