Cloning and expression of a 12 kDa serospecific epitope of Wuchereria bancrofti.
The immunoscreening of a microfilarial cDNA library of Wuchereria bancrofti with microfilaraemic sera revealed many positive clones expressing filarial antigens. One immunoreactive clone, designated PMR1, was shown to encode a protein of 114 amino acid residues. The cDNA fragment was subcloned into an expression vector, Pinpoint XaT. The resulting recombinant (r)PMR1-biotin fusion protein was expressed in Escherichia coli (BL21 [DE3] pLys) and was affinity purified on avidin resin. Analysis of sera of different groups for filarial antibodies against rPMR1 showed it to be highly reactive with microfilaraemic and clinical filarial sera compared to its reactivity with endemic and nonendemic controls. This indicates that the gene sequence of cDNA is expressing an immunodominant epitope, which could be useful in serodiagnosis of lymphatic filariasis.
Dabir
Pankaj
P
Department of Biochemistry, JB Tropical Disease Research Centre, Mahatma Gandhi Institute of Medical Sciences, Sevagram, Wardha, India.
Dabir
Snehal
S
Reddy
M V R
MV
D000818:Animals; D000947:Antigens, Helminth / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D001710:Biotin / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D003001:Cloning, Molecular; D004605:Elephantiasis, Filarial / Q000097:blood / Q000175:diagnosis / Q000276:immunology*; D015801:Helminth Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D006801:Humans; D016056:Immunodominant Epitopes / Q000235:genetics / Q000276:immunology*; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D014958:Wuchereria bancrofti / Q000235:genetics* / Q000276:immunology / Q000378:metabolism
D000947:Antigens, Helminth; D015801:Helminth Proteins; D016056:Immunodominant Epitopes; C525261:PMR1 protein, Wuchereria bancrofti; D011993:Recombinant Fusion Proteins; D001710:Biotin