Expression of human metallothionein-II fusion protein in Escherichia coli.
In order to obtain antibody against metallothionein-II (MT-II), a capsid protein metallothionein fusion protein was prepared. A gene encoding human MT-II was cloned into a plasmid for expression of MT fusion protein in Escherichia coli. MT cDNA was generated from human astrocytoma U373MG and amplified by polymerase chain reaction. The nucleotide sequence was identical to reported MT-II. The cDNA was inserted into plasmid pGEM EXTM-2 which carries the T7 promoter and T7 phage 10A major head protein. This expressed phage protein-MT fusion protein, has a molecular weight of 37kDa, forms inclusion bodies and constitutes about 20% of the total protein in transformed E. coli.
Yamazaki
S
S
Department of Hospital Pharmacy, Jichi Medical School, Tochigi, Japan.
Nakanishi
M
M
Hamamoto
T
T
Hirata
H
H
Ebihara
A
A
Tokue
A
A
Kagawa
Y
Y
D000595:Amino Acid Sequence; D017123:Bacteriophage T7 / Q000235:genetics; D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli; D006801:Humans; D008668:Metallothionein / Q000235:genetics*; D008969:Molecular Sequence Data; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000235:genetics; D014407:Tumor Cells, Cultured
D011993:Recombinant Fusion Proteins; D004247:DNA; D008668:Metallothionein