轮状病毒VP7-U基因在胡萝卜中表达的免疫原性分析  

Immunogenicity analysis of rotavirus VP7-U gene expression in carrots

作　　者：刘满杏[1,2] 李霞[2] 刘荣堂[1] 杨慧[2] 赵丽[2] 李鹏[2] 

机构地区：[1]甘肃农业大学生命科学技术学院,甘肃兰州730070 [2]北京市农林科学院蔬菜研究中心,北京100097

出　　处：《草业科学》2012年第12期1827-1831,共5页Pratacultural Science

基　　金：国家863专项(2007AA100505)

摘　　要：轮状病毒是一种危害人类健康的腹泻病毒,缺乏可控性药物。将优化部分密码子的轮状病毒VP7基因(VP7-U基因)整合到胡萝卜(Daucus carota)植株中,并利用PCR、Western blot、ELISA等方法检测转VP7-U基因胡萝卜的蛋白表达情况。然后利用不同浓度的阳性VP7-U胡萝卜蛋白灌服小鼠,以灌服正常胡萝卜植株蛋白的小鼠为阴性对照,以灌服阳性细菌蛋白的小鼠为阳性对照,灌服4次后利用ELISA方法检测小鼠体内的IgG含量。结果表明,转VP7-U基因的胡萝卜植株在蛋白水平上有目的抗原的表达并且其表达含量有一定的增加。灌服阳性胡萝卜蛋白的小鼠IgG含量高于灌服阴性胡萝卜蛋白的小鼠,但低于灌服阳性细菌蛋白的小鼠。然而统计分析表明,阳性小鼠同阴性小鼠体内的IgG含量差异不显著。转基因胡萝卜口服疫苗的发展还有很多困难,需进一步深入研究。Rotavirus is a diarrhea virus that is hazard to human health, and there are no effective drugs to control. In this study, firstly, some optimized codon of rotavirus VP7 gene （VP7-U gene） were integrated into carrot （Daucus carota） plants, to increase the amount of protein expression and then detect protein expression of modified gene of carrot by PCR, Western-blot, and ELISA methods. Secondly, using differ- ent concentrations of positive VPT-U carrots protein to drench mice, the mice that drenched normal carrot plant protein were taken as negative control, drenched positive bacterial proteins as positive control, and then detected the content of IgG in mice after 4 times＇ drenching by ELISA method. The results showed that the modified VP7-U genes of carrot plants had increased on target antigen expression and its levels. LgG content in mice of drenching positive proteins were higher than those with negative carrot proteins, while were lower than those with positive bacterial proteins. However, there was no significant difference for the IgG content in between positive control and negative control. There are many difficulties for the development of transgenie carrot oral vaccine, which need further study.

关 键 词：轮状病毒 转基因胡萝卜 VP7-U基因 口服疫苗 

分 类 号：Q943.2[生物学—植物学]