|
|
|
|
Prokaryotic expression and polyclonal antibody preparation of macrophage
migration inhibitory factor of Trichomonas vaginalis |
| LU Ye1, ZHU Yu-lian1， ZHU Li1, LIU Qing1， ZHU Yu-rong1, LING Wei1, TANG Chang-xia1 |
| (1. School of Medicine, Jiangsu University, Zhenjiang Jiangsu 212013; 2. National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention, Shanghai 200025， China）
|
|
|
|
|
Abstract Objective: To prokaryotically express the macrophage migration inhibitory factor of Trichomonas vaginalis(TvMIF) and produce its polyclonal antibody in rabbits. Methods: The TvMIF gene was amplified by PCR, and cloned into pTG19T vector.Then positive gene fragment was inserted into pET30a(+) vector after doubledigesting and sequencing.The recombinant plasmid was transformed into E.coli strain BL21(DE3), and the recombinant protein TvMIF was inducibly expressed with IPTG and purified by NiNAT affinity chromatography and then immunized rabbits. The polyclonal antibody was identified by Western blotting. Results: The TvMIF gene was amplified from Trichomonas vaginalis cDNA by PCR. Squencing analysis showed that its sequence is the same as that in GenBank. The recombinant plasmid of pET30aTvMIF was constructed, and the molecular weight of the recombinant TvMIF protein was about 15.5 kDa,which was in the form of soluble protein. The polyclonal antibody of TvMIF recombinant protein was obtained, and the specific band of TvMIF was detected by Western blotting. Conclusion: This study successfully purified the MIF protein of Trichomonas vaginalis and prepared its polyclonal antibody, which will facilitate the research on the pathogenic mechanism of Trichomonas vaginalis.
|
|
Received: 28 June 2018
|
|
|
|
| ［1］Salawu OT, Esume CN. Frequency of human immunodeficiency virus (HIV) in Trichomonas vaginalis infected women in Badagry, Lagos, Nigeria\[J\]. J Reprod Infertil, 2016, 17(1):61-63.［2］Kojic EM. Human papillomavirus(HPV) and trichomonas: common, concerning, and challenging sexually transmitted infections\[J\]. Med Health R I, 2012, 95(8):255-257.［3］Hosny AEMS, ElKhayat W, Kashef MT, et al. Association between preterm labor and genitourinary tract infections caused by Trichomonas vaginalis, Mycoplasma hominis, Gramnegative bacilli, and coryneforms\[J\]. J Chin Med Assoc, 2017, 80(9):575-581.［4］王冬冬, 杨真勇, 李笑南,等. 阴道毛滴虫病及其与生殖器恶性肿瘤相关性的研究进展\[J\].中国微生态学杂志, 2017, 29(8):962-965.［5］Calandra T, Bucala R. Macrophage migration inhibitory factor (MIF): a glucocorticoid counterregulator within the immune system\[J\]. Crit Rev Immunol, 1997, 17(1):77-88.［6］张梓岗, 陈小美, 苏丹华,等. 刚地弓形虫硫氧还蛋白基因的克隆表达及鉴定\[J\]. 中国血吸虫病防治杂志, 2016, 28(3):289-292.［7］Lang T, Foote A, Lee JP, et al. MIF: implications in the pathoetiology of systemic lupus erythematosus\[J\]. Front Immunol, 2015, 6:577.［8］Fukaya R, Ohta S, Yaguchi T, et al. MIF maintains the tumorigenic capacity of brain tumorinitiating cells by directly inhibiting p53\[J\]. Cancer Res, 2016, 76(9):2813-2823.［9］Brock SE, Rendon BE, Xin D, et al. MIF family members cooperatively inhibit p53 expression and activity\[J\]. PLoS One, 2014, 9(6): e99795.［10］Han C,Lin Y,Shan G,et al. Plasma concentration of malaria parasitederived macrophage migration inhibitory factor in uncomplicated malaria patients correlates with parasitemia and disease severity\[J\]. Clin Vaccine Immunol, 2010, 17(10): 1524-1532.［11］Bennuru S, Semnani R, Meng Z, et al. Brugia malayi excreted/secreted proteins at the host/parasite interface: stage and genderspecific proteomic profiling\[J\]. PLoS Negl Trop Dis, 2009, 3(4): e410. |
|
|
|
2019, Vol. 29 
