[Screening of T7 phage-display cDNA library from liver of Microtus fortis with extracts of schistosomulum and characterization of the positive clones].
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.
Jia
Renchu
R
National Laboratory of Animal Schitosomiasis Control, Key Laboratory of Animal Parasitology, Ministry of Agriculture, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200232, China.
Sun
Yi
Y
Liu
Jinming
J
Fu
Zhiqiang
Z
Yuan
Chunxiu
C
Shi
Yaojun
Y
Lu
Ke
K
Sun
Huan
H
Li
Hao
H
Jin
Yamei
Y
Lin
Jiaojiao
J
D000818:Animals; D003411:Arvicolinae / Q000235:genetics* / Q000469:parasitology; D017123:Bacteriophage T7 / Q000235:genetics; D003001:Cloning, Molecular; D020224:Expressed Sequence Tags; D015723:Gene Library; D017238:Genes, Helminth / Q000235:genetics; D007113:Immunity, Innate / Q000235:genetics*; D007814:Larva / Q000235:genetics / Q000254:growth & development; D008099:Liver / Q000737:chemistry*; D012549:Schistosoma japonicum / Q000235:genetics* / Q000254:growth & development