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Published on: March 24, 2017
[Preparation and identification of polyclonal antibody to serotype I Mareks disease virus sorf2 protein]
Ning Cui1, Shuai Su, Jiuqing Li
1Animal Science and Technology College, Shandong Agricultural University, Tai' an 271018, China. cnydia@163.com
Objective:
To obtain mice and rabbit polyclonal antibody of serotype I Marek's disease virus (MDV) sorf2 protein with higher titer and to identify the specificity.
Methods:
Using serotype I MDV GX0101 as template, we amplified sorf2 gene and then cloned it into pET-28a (+) and pET-32a (+) respectively. The recombinant plasmid pET-28a-sorf2 and pET-32a-sorf2 was separately transformed into Escherichia coli BL21 (Rosetta) competent cell which was induced with isopropylthio-beta-D-galactoside (IPTG). After purification, immuned 6-8 Balb/c mice and adult New Zealand white rabbit with the purified fusion protein and the anti-sorf2 polyclonal antibody were prepared. The specificity of the serum was detected by Western blot and the indirect immunofluorescence assay (IFA) method.
Results:
Serotype I MDV sorf2 protein was expressed successfully in the recombinant E coli. Mice and rabbit anti-sorf2 polyclonal antibody with higher titer could react with sorf2 protein specifically.
Conclusion:
The prepared anti-sorf2 polyclonal antibody could identify MDV sorf2 gene deletion strain specifically. In addition, it could be used for differential of MDV vaccine poison HVT and serotype I MDV, which was useful for the separation and identification of clinical MDV.
