SYNTHESIS OF THE BIOLOGICALLY-ACTIVE BETA-SUBUNIT OF HUMAN NERVE GROWTH-FACTOR IN ESCHERICHIA-COLI

被引：15

作者：

NEGRO, A ^{[1
]}

MARTINI, I ^{[1
]}

BIGON, E ^{[1
]}

CAZZOLA, F ^{[1
]}

MINOZZI, C ^{[1
]}

SKAPER, SD ^{[1
]}

CALLEGARO, L ^{[1
]}

机构：

[1] FIDIA RES LABS, RES LABS, I-35031 ABANO TERME, ITALY

来源：

GENE | 1992年 / 110卷 / 02期

关键词：

RECOMBINANT DNA; PLASMID EXPRESSION VECTOR; PHAGE-LAMBDA PR AND PL PROMOTERS; IMMUNOLOGICAL CROSS-REACTIVITY; HOMODIMER; DORSAL ROOT GANGLION; NEURITE OUTGROWTH;

D O I：

10.1016/0378-1119(92)90657-B

中图分类号：

Q3 [遗传学];

学科分类号：

071007 ; 090102 ;

摘要：

The gene (NGFB) encoding the beta subunit of mature human nerve growth factor (hNGFB) was subcloned into the pJLA503 expression vector under the control of bacteriophage promoters P(R) and P(L), and expressed in Escherichia coli. The recombinant protein represented approximately 3% of the total cellular protein. Biologically active hNGFB was solubilized (0.2% total NGFB) and purified by cation-exchange chromatography and it yielded two bands on polyacrylamide-gel electrophoresis under nonreducing conditions, corresponding to the monomeric (14 kDa) and homodimeric (26.5 kDa) forms of the molecule. Both hNGFB forms were immunopositive on Western blots with rabbit anti-NGFB antibodies; however, following additional purification, only the species corresponding to the hNGFB homodimer was biologically active on cultured chicken dorsal root ganglion neurons. These results demonstrate the feasibility of synthesizing the biologically active form of hNGFB in E. coli.

引用

页码：251 / 256

页数：6

共 22 条

[1] NERVE GROWTH FACTOR FROM MOUSE SUBMAXILLARY GLAND - AMINO ACID SEQUENCE [J].