Expression and polyclonal antibody preparation of the tegument protein UL48 encoded by MDV

VernacularTitle:马立克氏病毒被膜蛋白VP16(UL48)的表达及多克隆抗体的制备
Author: Jingjing SONG ; Chen DENG ; Shanli WU ; Hainan ZHENG ; Peifeng YU ; Mengyun WANG ; Xiaolu ZHOU ; Yujing ZHANG ; Yongxing AI
Keywords: Marek's disease virus; UL48; polyclonal antibody; intein; thrombin
From: Chinese Journal of Veterinary Science 2017;37(8):1473-1478
CountryChina
Language:Chinese
Abstract: UL48 plays essential role in replication of MDV genome and interacts with UL36 as well as other MDV tegument proteins.To investigate the interaction between UL48 and UL36 during MDV oncogenisis,antibody against UL48 was prepared and characterized in current study.UL48 gene was amplified from MDV-Ⅰ genome and then subcloned into pTYB1 and pGEX-4T3 vectors for UL48 expression with induction of IPTG in BL21(DE3) E..coli cells.Chitin-sepharose and Glutathion-sepharose were,respectively,used to purify fusion protein intein-UL48 and GST-UL48.Four subcutaneous injections of intein-UL48 fusion protein were done on the lower back and the thigh of rabbit and then other three injections with an interval 10 days.The titer of antibody was measured by the sandwich ELISA with UL48 protein isolated from GST-UL48 after cleavage of thrombin.Western blot was carried out for specificity analysis of antibody against UL48 protein.The results suggested that UL48 antibody was succesfully prepared,and its titer was 1 ∶ 512 000.