Construction, expression, and identification of Luffin P1-KDEL
LIU Shulei
HE Wei
WANG Rupeng
WU Jun
HU Xiaoheng
Abstract：Objective Luffin P1 as a new member of luffin family is an effective ribosome inactivating protein (RIP).In this study, we aimed to construct prokaryotic expression plasmid PET32a (+)-Luffin P1 for expressing, purifying,and identifying the expression protein Luffin P1.We subcloned a recombinant LuffinPl DNA fragment into an expression plasmid pET32a(+), and identified the recombined plas-mid by restriction enzyme digestion and DNA sequencing.After the recombinant plasmid was transformed into bacteria BL21(DE3), the ex-pression of fusion protein Luffin P1 was induced by IPTG and identified by Western blotting using anti-His Ab, then the fusion protein was purified, digested by enterokinase, and re-purified.The new recombinant plasmid pET32a (+)-Luffin P1 was successfully constructed, and the correct fusion protein of Luffin P1 was highly purified.These results imply that Luffin P1 protein cand be acquired through genetic en-gineering, which provides a experimental basis for function research of Luffin P1 and production of immunotoxin warheads.
Keywords：ImmunotoxinRibosome-inactivating proteinLuffin P1
Publication Date：2010-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：5( 16-20 )