[Purification and biological activity of rh-leptin expressed in Escherichia coli].
The human leptin was successfully expressed with high level in E. coli under the control of PL promotor. The yield of recombinant protein was over 40% of total cellular protein and expressed as inclusion bodies. The recombinant human leptin (rh-leptin) was purified with gel filtration, anion-exchange and reverse chromatography. Refolding was achieved by gradually reducing denaturant using a diafiltration method. The refolded rh-leptin was characterized by SDS-PAGE, Western-blotting and its first 15 amino acid residues sequence of the N-terminal. The purified product was found to be biologically active, reducing the food intake and body weight gain upon testing in BALB/c mice.
Zhao
Y R
YR
Shandong Tumor Biotherapy Centre, Institute of Basic Medicine, Shandong Academy of Medical Sciences, Jinan 250062, China. yrzhao@public.jn.sd.ch
Wang
J F
JF
You
L
L
Gao
C Y
CY
Tian
Z G
ZG
Zhang
J
J
Han
N
N
Yin
J
J
Sun
R
R
D000818:Animals; D004435:Eating / Q000187:drug effects; D004926:Escherichia coli / Q000235:genetics*; D005260:Female; D006801:Humans; D020738:Leptin / Q000096:biosynthesis* / Q000302:isolation & purification / Q000494:pharmacology; D051379:Mice; D008807:Mice, Inbred BALB C; D011994:Recombinant Proteins / Q000096:biosynthesis*; D015430:Weight Gain / Q000187:drug effects
D020738:Leptin; D011994:Recombinant Proteins