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Indian Journal of Biotechnology (IJBT) >
IJBT Vol.04 [2005] >
IJBT Vol.04(3) [July 2005] >
| Title: | Cloning, characterization and expression of bovine (Bos indicus) tumour necrosis factor- |
| Authors: | Gupta, Praveen K
Bind, R B
Walunj, Sameer S
Saini, Mohini |
| Keywords: | Bos indicus
cDNA sequence
expression
TNF- |
| Issue Date: | Jul-2005 |
| Publisher: | CSIR |
| IPC Code: | Int. Cl.7 C12N15/10, 15/28 |
| Abstract: | The gene for tumour necrosis factor-alpha
(TNF- ) was amplified from cDNA pool prepared from LPS-stimulated bovine
peripheral blood mononuclear cells isolated from Indian cattle. The amplified
TNF- gene was cloned and nucleotide sequences were determined. Homology
comparison of nucleotide and predicted amino acid sequences revealed similarity
at nucleotide and amino acid level in both exotic cattle and goat. The coding
sequence of mature TNF- (without its signal sequence and transmembrane anchor) was
expressed as fusion protein with N-terminal polyhistidine using prokaryotic
expression vector. The expressed protein was present as insoluble inclusion
bodies. The recombinant protein was solubilized with urea and purified using
Ni-agarose affinity chromatography. The purified recombinant TNF- was
characterized in SDS-PAGE and in western blotting. |
| Page(s): | 363-366 |
| ISSN: | 0975-0967 (Online); 0972-5849 (Print) |
| Source: | IJBT Vol.04(3) [July 2005]
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