Expression and identification of the ADF-linker-3-1E gene of Eimeria acervulina of XXXX .

@entity6874 @entity3208 @entity19 @entity1573 @entity2975 @entity214 @entity526
UNASSIGNED: Coccidiosis is a widely distributed disease with higher mortality and morbidity, which is caused by several species of protozoan parasites belonging to the genus Eimeria and recognized as a serious challenge for the poultry industry. This research was conducted to construct the recombinant plasmid pET32a(+)-ADF-linker-3-1E of Eimeria acervulina (E. acervulina) of the @entity1573 and test the bioactivity of the ADF-linker-3-1E protein. The ADF-linker-3-1E gene of E. acervulina of the @entity1573 was cloned by splicing by overlap extension by the polymerase chain reaction (SOE-PCR) and then inserted into the pET32a(+) to construct the recombinant plasmid pET32a(+)-ADF-linker-3-1E. The recombinant plasmid was transformed into @entity526 Rosetta (DE3) competent cells and then induced by @entity6874 (0.6  mmol/L). The expressed product in the culture medium was identified by the @entity3208 - @entity2975 gel electrophoresis ( @entity214 -PAGE). The bioactivity of the ADF-linker-3-1E protein was tested by Western blotting. The result showed that the amplified ADF-linker-3-1E gene was about 1346  bp. The PCR amplification with the recombinant plasmid pET-32a(+)-ADF-linker-3-1E as a template resulted in a special band of 1346  bp. The digested products resulted in two fragments of 1346  bp target fragment and 5.9  kb pET-32a(+)-vector fragment. The results indicated that the ADF-linker3-1E gene was successfully inserted into the pET-32a(+)-vector. The expressed products in the culture medium resulted in a single band of approximately 54.8  kDa by @entity214 -PAGE. Western blotting analysis indicated that the recombinant protein could be reacted specifically with His-Tag(2A8) @entity19 mAb. This study indicated that the ADF-linker-3-1E protein with good bioactivity was successfully obtained, which laid a foundation for the exploitation of the nuclear vaccine by using the ADF-linker-3-1E protein.
