Abstract
We have previously developed the TraT display system to express the preS1 peptide of human hepatitis B virus (HBV) and the snake venom rhodostomin (RHO) on the surface of Escherichia coli. In this study, we modified the pT2 vector by adding a thrombin cutting site and a phosphorylation tag of protein kinase A before the multiple restriction enzyme sites. The modified vector allowed us to label the TraT fusion protein (TraT-RHO) with [32P] and to increase the detection sensitivity of TraT-RHO expression bacteria binding to and being internalized into BHK-21 cells. After the thrombin cleavage, the isotope labeled RHO could be detected in a free form. We therefore suggest that the new version of pT2 vector, pT2-KL, will facilitate to identify the counterpart of displayed peptide.
Original language | English |
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Pages (from-to) | 115-122 |
Number of pages | 8 |
Journal | Journal of Biotechnology |
Volume | 78 |
Issue number | 2 |
DOIs | |
State | Published - 10 03 2000 |
Externally published | Yes |
Keywords
- Bacterial display
- Protein kinase A
- Rhodostomin
- Thrombin
- TraT lipoprotein