Yamazaki S, Nakanishi M, Hamamoto T, Hirata H, Ebihara A, Tokue A, Kagawa Y
Department of Hospital Pharmacy, Jichi Medical School, Tochigi, Japan.
Biochem Int. 1992 Nov;28(3):451-60.
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.
为了获得抗金属硫蛋白-II（MT-II）的抗体，制备了一种衣壳蛋白金属硫蛋白融合蛋白。将编码人MT-II的基因克隆到质粒中，以便在大肠杆菌中表达MT融合蛋白。MT cDNA是从人星形细胞瘤U373MG中产生的，并通过聚合酶链反应进行扩增。核苷酸序列与报道的MT-II相同。将该cDNA插入携带T7启动子和T7噬菌体10A主要头部蛋白的质粒pGEM EXTM-2中。这种表达的噬菌体蛋白-MT融合蛋白分子量为37kDa，形成包涵体，在转化的大肠杆菌中占总蛋白的约20%。
