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Expressing the immunodominant projection domain of infectious bursal disease virus fused to the fragment crystallizable of chicken IgY in yellow maize for a prospective edible vaccine.
Molecular Immunology ( IF 3.6 ) Pub Date : 2019-12-24 , DOI: 10.1016/j.molimm.2019.12.015
Reda Salem 1 , Shireen K Assem 1 , Omar A Omar 1 , Ahmed A Khalil 2 , Mahmoud A Basry 1 , Fatma R Waly 1 , Noha Samir 1 , Alaa A El-Kholy 2
Affiliation  

Control of Infectious bursal disease virus (IBDV) in endemic countries has been based on early immunization of chicks using conventional live or inactivated vaccines that became not fully effectual and have biosafety concerns. This endeavor seeks generating a recombinant chimeric protein merging the projection domain (PD) of IBDV VP2 capsid with the fragment crystallizable (Fc) of avian IgY (FcIgY), in maize as a prospective poultry edible vaccine. The PD sequence was built on the basis of very virulent IBDV isolates circulating in Egypt. After optimization of codon-usage in maize, sequences of PD and FcIgY were effectively expressed in two elites of yellow maize via bombardment transformation in immature embryos. Chimeric protein amount in stable transgenic samples ranged from1.36% to 3.03% of the total soluble protein based on tissue age and maize cultivar. IBDV VP2 coding sequence was amplified from viral RNA, cloned, and expressed in E. coli. A group of Balb/C mice were hyper-immunized with purified recombinant VP2 protein for raising anti- recombinant VP2 antibodies (anti-rVP2 Ab). Proper expression in maize and immunoreactivity of the chimeric protein (PD-FcIgY) to chicken anti- IBDV and anti-rVP2 Ab were confirmed by both direct and indirect double antibody sandwich (DAS)-ELISAs as well as western blotting. Seeds of regenerated transgenic maize will be validated for chickens as edible vaccination in further studies.

中文翻译:

表达与黄色玉米中鸡IgY的可结晶片段融合的传染性法氏囊病病毒的免疫优势投射域,从而可用于未来的可食用疫苗。

在流行国家控制传染性法氏囊病病毒(IBDV)的基础是使用常规活疫苗或灭活疫苗对小鸡进行早期免疫,这种疫苗不能完全发挥作用,并具有生物安全性。该努力寻求在玉米中产生重组嵌合蛋白,其将IBDV VP2衣壳的投影结构域(PD)与禽IgY的可结晶片段(Fc)(FcIgY)融合,作为预期的家禽可食用疫苗。PD序列是基于在埃及传播的极强毒力的IBDV分离株构建的。优化玉米密码子使用后,PD和FcIgY序列通过未成熟胚中的轰击转化在两个黄玉米精英中有效表达。稳定的转基因样品中的嵌合蛋白含量范围为1.36%至3。基于组织年龄和玉米品种的总可溶性蛋白的03%。从病毒RNA扩增IBDV VP2编码序列,克隆并在大肠杆菌中表达。用纯化的重组VP2蛋白高度免疫一组Balb / C小鼠,以产生抗重组VP2抗体(抗rVP2Ab)。通过直接和间接双抗体夹心(DAS)-ELISA以及Western印迹证实了嵌合蛋白(PD-FcIgY)在鸡中的正确表达以及对鸡抗IBDV和抗rVP2 Ab的免疫反应性。再生的转基因玉米种子将在食用鸡疫苗中得到进一步研究的验证。用纯化的重组VP2蛋白高度免疫一组Balb / C小鼠,以产生抗重组VP2抗体(抗rVP2Ab)。通过直接和间接双抗体夹心(DAS)-ELISA以及Western印迹证实了嵌合蛋白(PD-FcIgY)在鸡中的正确表达以及对鸡抗IBDV和抗rVP2 Ab的免疫反应性。再生的转基因玉米种子将在食用鸡疫苗中得到进一步研究的验证。用纯化的重组VP2蛋白高度免疫一组Balb / C小鼠,以产生抗重组VP2抗体(抗rVP2Ab)。通过直接和间接双抗体夹心(DAS)-ELISA以及Western印迹证实了嵌合蛋白(PD-FcIgY)在鸡中的正确表达以及对鸡抗IBDV和抗rVP2 Ab的免疫反应性。再生的转基因玉米种子将在食用鸡疫苗中得到进一步研究的验证。
更新日期:2019-12-27
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