Screening of T7 phage-display cDNA library from liver of Microtus fortis with extracts of schistosomulum and characterization of the positive clones / 生物工程学报

ABSTRACT

Microtus fortis is naturally resisitent to Schistosoma japonicum. In order to find schistosome-resistence-related genes of Microtus fortis, a T7 phage-display cDNA library from liver of Microtus fortis was screened with the soluble lysate of schistosomula. The specific phages were enriched 375-fold after 3 rounds of biopanning. Ninety-two positive clones picked at random were sequenced and 19 ESTs including 6 unreported genes were obtained. Compared with the negative phage clone control, five positive clones, No.4 (GenBank Accession No.: EW968294), No.13 (GenBank Accession No.: EW968303), No.14 (GenBank Accession No.: EW968304), No.15 (GenBank Accession No.: EW968305) and No.18 (GenBank Accession No.: EW968308) could induce significantly higher schistosomula mortality rate when co-cultivated with schistosomula. According to the function analysis and the shistosomula-killing effect in vitro, the genes encoding CASP8 and FADD-like apoptosis regulator isoform protein, alpha-2-HS-glycoprotein, M4 protein, R3H domain (binds single-stranded nucleic acids) isoform 2 and 3 previously unreported proteins (No.14, No.15 and No.18) obtained here, were schistosomiasis-resistence-related genes of Microtus fortis.