Serial recombinant expression and anti-tumor activity in vitro of antibiotic peptide Alloferon-1

• Published in: Zhejiang da xue xue bao. Journal of Zhejiang University. Medical sciences. Yi xue ban Vol. 40; no. 5; pp. 501 - 507
• Main Authors: Xu, Xiao-yin, Yan, Jie, Sun, Qi
• Format: Journal Article
• Language: Chinese
• Published: China 01.09.2011
• Subjects: Animals; Antineoplastic Agents - pharmacology; Cell Line, Tumor; Genetic Vectors; Humans; Male; Mice; Mice, Inbred BALB C; Peptides - genetics; Peptides - pharmacology; Recombinant Fusion Proteins - biosynthesis; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - pharmacology
• ISSN: 1008-9292

To generate a recombinant expression system of repeated serial antibiotic peptide Alloferon-1 DNA segment with trypsin digestion site and to determine its anti-tumor activity in vitro. A 14 repeated serial DNA segment of Alloferon-1 with a lysine residual at the C-end that acts as the trypsin digestion site was constructed. pET42a vector and E.coli BL21DE3 were applied to generate the prokaryotic expression system of the repeated serial DNA segment of Alloferon-1. The yield of target recombinant product was measured by SDS-PAGE and Bio-Rad Gel image system. Ni-NTA affinity column, trypsin digestion and Sephadex G-50 column were used to purify 14 rAlloferon-1-K fusion protein and rAlloferon-1-K monomer. By using the co-cultivation of BALB/c mouse splenocyte with K562, KB or SGC tumor cells and CCK-8 detection method, the effects of rAlloferon-1-K, chemosynthetic Alloferon-1 (cAlloferon-1) and Alloferon-1-K (cAlloferon-1-K) on the growth and proliferation of tumor cells were detected. The prokaryotic expression