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Indian Journal of Biotechnology (IJBT) >
IJBT Vol.04 [2005] >
IJBT Vol.04(1) [January 2005] >
| Title: | Cloning and expression of bovine (Bos indicus) interleukin-2 in Escherichia coli |
| Authors: | Patil, Amar A
Saini, Mohini
Sharma, Sameer
Bind, R B
Gupta, Praveen K |
| Keywords: | Bos indicus
cDNA sequence
expression
interleukin-2
prokaryotic
recombinant |
| Issue Date: | Jan-2005 |
| Publisher: | CSIR |
| IPC Code: | Int. Cl.7 C 12 N 15/26 |
| Abstract: | The gene for interleukin-2 (IL-2) was amplified
from cDNA pool prepared from ConA-stimulated peripheral blood mononuclear cells
(PBMCs) isolated from Indian cattle (Bos
indicus). The amplified IL-2 gene was cloned and nucleotide sequences were
determined. Homology comparison of nucleotide and predicted amino acid
sequences revealed similarity of sequence and conservation of crucial amino acids
with exotic cattle (B. taurus). The
coding sequence of IL-2 (without its own signal sequence) was subsequently
expressed as fusion protein with polyhistidine fusion tag in Escherichia coli, 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 IL-2 was
characterized in SDS-PAGE and in western blotting. |
| Page(s): | 88-92 |
| ISSN: | 0975-0967 (Online); 0972-5849 (Print) |
| Source: | IJBT Vol.04(1) [January 2005]
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