Construction and immunoscreening of cDNA library of Babesia microti

• DOI: 10.3760/cma.j.issn.1673-4122.2013.03.002
• Publication Date: 2013-05-28
• Authors: Jiahui Sun; Bin Xu; Chuan Ju; Jun‐Hu Chen; Yan Lu; Shaohong Chen; Jian-ping Han
• Keywords: Immunoscreening

Objective To construct a cDNA library of Babesia microti and immunoscreen candidate antigens for immuno-diagnosis infection.Methods The blood samples were collected from the mice infected with B.microti 7 days post infection when parasitemia density reached 70％.Total RNA was extracted mRNA obtained by purifying kit used to cDNA-library.The immunoscreened pooled sera obtain positive clones.The inserted fragments clones identified PCR amplification,and genes sequenced analyzed their homology.Results titer 5.2 × 101 plaque forming unit (pfu)/ml.The fragment length ranged 500 3 000 bp recombination efficiency being 91.0％.Six clones,Bm2,Bm4,Bm6,Bm7,Bm9 Bm15 found about 633,614,1 073,890,1 001 1 032 bp,respectively.Sequence analysis revealed that all 6 contained open reading frames.The deduced amino acid sequences 159,100,254,217,60 244 residues,with Mr 17 900,11 400,28 600,24 000,6 800 28 900,respectively.Conclusion A has been constructed identified,which can be recognized B.microti.This study provided preliminary information further identification highly reactive development immunodiagnostics infection.

Key words: Babesia ; Immunoscreening