Gene expression and function study of fusion immunotoxin anti-Her-2-scFv-SEC2 in Escherichia coli.
The whole sec2 DNA fragment was obtained by polymerase chain reaction with total genomic DNA extracted from Staphylococcus aureus. The newly discovered gene contains 717 bp (GenBank Accession number AY450554) and encodes 239 amino acids in accordance with the protein sequence reported in GenBank. The sec2 gene was fused to anti-Her-2 scFv gene with a DNA linker at the upstream to construct the fusion gene of immunotoxin which was subcloned to expression vector pET-32a(+) and expressed efficiently in Escherichia coli. The purified fusion immunotoxin could target the HER-2 overexpressed by breast tumor cell SK-Br-3 in vitro and demonstrate tumor-inhibition effect on SK-Br-3.
Ming-Kai
Xu
X
Institute of Applied Ecology, Chinese Academy of Science, Shanyang 110016, China. mingkaixu@yahoo.com.cn
Cheng-Gang
Zhang
Z
D000970:Antineoplastic Agents; D045744:Cell Line, Tumor; D004768:Enterotoxins / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D006801:Humans; D000922:Immunotoxins / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism; D008969:Molecular Sequence Data; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000276:immunology*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
D000970:Antineoplastic Agents; D004768:Enterotoxins; D000922:Immunotoxins; D018719:Erb-b2 Receptor Tyrosine Kinases; D011994:Recombinant Proteins; C034959:enterotoxin C, staphylococcal