Selection Of A Peptide Binding To Brush Border Membrane Vesicles From Nilaparvata Lugens With A Phage Display Peptide Library

One thousand of last-instar Nilaparvata lugens nymphs were dissected, guts from dissection were collected to yield a pool of brush border membrane vesicles(BBMV) after centrifugation. A peptide binding to BBMV was screened using Phage Display, an unbound peptide was also screened to use as a contrast. Sequences of peptides and green fluorescence protein (GFP) were combined by polymerase chain reaction (PCR). The recombined sequences were cloned into pMD18-T vector and pET-32a vector successfully, and the recombinant plasmids were transformed into E. coil JM109 and BL21 respectively. Under the induction of IPTG, a fusion protein with a molecular mass about 45kDa was well produced. Using His tag on pET-32a vector, the fusion protein was purified with a Ni column. The purified fusion protein was then fed to last-instar Nilaparvata lugens nymphs, guts were collected and rinsed after dissection and transferred to fluorescence light microscopy to observe the binding effect. Images showed that the fusion protein was bound well to the gut.