Expression of nine-banded armadillo (Dasypus novemcinctus) interleukin-2 in E. coli.
The nine-banded armadillo (Dasypus novemcinctus) is the only immunologically intact animal that regularly develops lepromatous-type leprosy when inoculated with Mycobacterium leprae. However, the ability to exploit this model for understanding the pathogenesis of leprosy has been limited by a lack of suitable immunological reagents. Recently, efforts began to sequence the entire armadillo genome, and this sequence information will help make possible the development of a wide array of new immunological reagents suitable for use with armadillos. Using the available sequence data, a region of high homology to interleukin-2 of other mammals was identified. Primers were designed to amplify the coding region corresponding to the mature peptide and its exact sequence was confirmed. cDNA was made from ConA-stimulated armadillo PBMC. The amplified coding region was sub-cloned into a pET expression vector and transformed into Escherichia coli for over-expression. The subsequent product was characterized by SDS-PAGE and bioassays. Tritiated thymidine incorporation by CTLL-2 and armadillo lymphoblasts confirmed functionality of the recombinant product. The advent of the D. novemcinctus genome sequence and subsequent generation of immunological tools will assist in advancing the armadillo as a translational model for leprosy.
Adams
J E
JE
Laboratory Research Branch National Hansen's Disease Programs, Louisiana State University, School of Veterinary Medicine, Skip Bertman Drive, Baton Rouge, LA 70803, USA.
Peña
M T
MT
Gillis
T P
TP
Williams
D L
DL
Adams
L B
LB
Truman
R W
RW
D000595:Amino Acid Sequence; D000818:Animals; D001135:Armadillos / Q000235:genetics* / Q000276:immunology; D002478:Cells, Cultured; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D016678:Genome; D006801:Humans; D007376:Interleukin-2 / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D017385:Sequence Homology
D007376:Interleukin-2; D011994:Recombinant Proteins