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Indian Journal of Experimental Biology (IJEB) >
IJEB Vol.49 [2011] >
IJEB Vol.49(09) [September 2011] >
| Title: | Cloning, protein expression and display of synthetic multi-epitope mycobacterial antigens on Salmonella typhi Ty21a cell surface |
| Authors: | Sarhan, Mohammed A A
Musa, Mustaffa
Zainuddin, Zainul F |
| Keywords: | Ice nucleation protein
Mycobacterium tuberculosis
Surface display
Ty21a |
| Issue Date: | Sep-2011 |
| Publisher: | NISCAIR-CSIR, India |
| Abstract: | Expressing proteins of interest as fusion to proteins of bacterial envelope
is a powerful technique for biotechnological and medical applications. The
synthetic gene (VacII) encoding for T-cell epitopes of selected genes of Mycobacterium tuberculosis namely,
ESAT6, MTP40, 38 kDa, and MPT64 was fused with N-
terminus of Pseudomonas syringae ice
nucleation protein (INP) outer membrane protein. The fused genes were
cloned into a bacterial expression vector pKK223-3. The recombinant protein was
purified by Ni-NAT column. VacII gene was displayed on the cell surface of Salmonella typhi Ty21a using N-terminal
region of ice nucleation proteins (INP) as an anchoring motif. Glycine method
confirmed that VacII was anchored on the cell surface. Western blot analysis
further identified the synthesis of INP derivatives containing the
N-terminal domain INP- VacII fusion protein of the expected size (52 kDa). |
| Page(s): | 645-653 |
| CC License: |  CC Attribution-Noncommercial-No Derivative Works 2.5 India |
| ISSN: | 0975-1009 (Online); 0019-5189 (Print) |
| Source: | IJEB Vol.49(09) [September 2011]
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