Prokaryotic expression and purification of human hepatic stimulator substance / 生理学报
Acta Physiologica Sinica
;
(6): 23-27, 2002.
Artigo
em Chinês
| WPRIM
| ID: wpr-272990
ABSTRACT
To explore the possibility of prokaryotic expression of human hepatic stimulator substance (hHSS), hHSS gene was inserted in the downstream of glutathion S-transferase (GST) in a pET-42a expression vector and recombinant GST-hHSS fusion protein was expressed under IPTG induction in BL-21(DE3) cells. The recombinant HSS was purified with His.Tag affinity chromatography, and its bioactivity was analyzed. The results showed that GST-hHSS fusion protein was expressed both as a soluble or a inclusive body in bacterial cytosol. The soluble GST-hHSS expression reached up to 30% of the whole soluble protein of bacteria as determined by densitometry. The cleavage of GST-hHSS fusion protein with Factor Xa produced two fragments of the protein, which sized 33 and 15 kD, respectively. The molecular weight of recombinant HSS protein was identical to theoretical deduction based on the DNA sequences. The protein homology of 15 kD hHSS could be efficiently eluted out after Factor Xa cleavage. It is further indicated that the recombinant hHSS is able to proliferate hepatoma cells of BEL-7402 in the preliminary experiments.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Peptídeos
/
Farmacologia
/
Proteínas Recombinantes de Fusão
/
Células Tumorais Cultivadas
/
Expressão Gênica
/
Divisão Celular
/
Substâncias de Crescimento
/
Genética
/
Glutationa Transferase
Limite:
Humanos
Idioma:
Chinês
Revista:
Acta Physiologica Sinica
Ano de publicação:
2002
Tipo de documento:
Artigo
Similares
MEDLINE
...
LILACS
LIS