1.The Second Clinical Medical College, Wenzhou Medical University, Wenzhou 325035, China; 2.Department of Surgical Oncology, Wenzhou People’s Hospital, Wenzhou 325000, China; 3.School of Basic Medical Sciences, Wenzhou Medical University, Wenzhou 325035, China.
CHEN Bi,XIE Shangdan,XIANG Chaoli, et al. Preparation and identification of human cytomegalovirus US31 protein specific antibody[J]. JOURNAL OF WEZHOU MEDICAL UNIVERSITY, 2019, 49(8): 547-551.
Abstract：Objective: To explore the preparation and identification of human cytomegalovirus (HCMV) US31 protein and its polyclonal antibody. Methods: After HCMV US31 gene was optimized by prokaryotic codon for whole gene synthesis and cloned to pET21a (+) prokaryotic expression vector, pET21a (+)/HCMV US31 recombinant plasmid was constructed, with identified sequence. Recombinant plasmid was transformed into Escherichia coli BL21 (DE3), target protein expression induced by IPTG, and US31 protein purified by Ni-NTA affinity chromatography. Immunized New Zealand rabbits were used to prepare polyclonal antibodies, and antibody titer and specificity was detected by ELISA, Western blot and immunofluorescence method. Results: High expression of HCMV US31 fusion protein was induced in Escherichia coli, and high purity target protein was obtained by nickel column affinity purification, and the specific IgG antibody was induced by immune New Zealand rabbits, with the valence being 1:49 600. ELISA, Western blot and immunofluorescence have confirmed that the prepared specific antibodies were able to identify US31 proteins expressed in cells. Conclusion: HCMV US31 protein and specific antibodies were successfully prepared, which laid foundations for further study of the biological function of US31 protein.
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