Syncytin, the envelope protein of human endogenous retrovirus family W, is expressed in specific tissues of healthy people and is generally believed to play a pivotal role in placenta development. To generate and identify the rabbit polyclonal antibodies against human syncytin, syncytin gene was applied by PCR and inserted into cloning vector. The recombinants were digested by BamH I/Not I, cloned into expression vector pET30a (+) and then transferred into E. coli BL21 (DE3). Fusion protein His/syncytin was induced by IPTG, and then analyzed by SDS-PAGE. The protein straps about 42kDa were cut, grinded and used to immunize New Zealand rabbits. Sera of the immunized rabbits were collected 10 days after the second enhanced immunization. The antiserum was detected by ELISA, Western blot and immunohistochemistry staining. The prokaryotic expressed pET30a (+)/syncytin vector was successfully constructed and fusion protein syncytin-His was expressed efficiently. The polyclonal antibodies were raised in the rabbits. The syncytin polyclonal antibodies reveal high titer and specificity. It may be applied in syncytin function research. In the second part of study, expression of syncytin in some malignant cells (10 leukemia or lymphoma cell lines and 30 peripheral blood samples of leukemia or lymphoma patients) was investigated. Transcripts of this gene were relatively quantified by real-time RT-PCR method, and its translated products were revealed via an indirect immunofluorescence assay. Results derived from both methods clearly demonstrated that syncytin was expressed in these malignant cells. Neither transcribed nor translated syncytin has been observed in healthy people’s blood cells. These observations suggest that expression of syncytin might be an indicator in diagnosis of leukemia disease. Further studies are warranted to clarify the relationship between abnormal expression of syncytin and leukemogenesis.
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