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朱进, 张云, 唐家琪, 李先富, 郭恒彬, 李喜文, 李彦舫. 转基因轮状病毒疫苗植物表达载体的构建及GUS基因表达[J]. 中国公共卫生, 2001, 17(10): 873-874. DOI: 10.11847/zgggws2001-17-10-09
引用本文: 朱进, 张云, 唐家琪, 李先富, 郭恒彬, 李喜文, 李彦舫. 转基因轮状病毒疫苗植物表达载体的构建及GUS基因表达[J]. 中国公共卫生, 2001, 17(10): 873-874. DOI: 10.11847/zgggws2001-17-10-09
ZHU Jin, . Construction of Rotavirus Vaccine Plant Cell Expression Vector and Expression of GUS Gene[J]. Chinese Journal of Public Health, 2001, 17(10): 873-874. DOI: 10.11847/zgggws2001-17-10-09
Citation: ZHU Jin, . Construction of Rotavirus Vaccine Plant Cell Expression Vector and Expression of GUS Gene[J]. Chinese Journal of Public Health, 2001, 17(10): 873-874. DOI: 10.11847/zgggws2001-17-10-09

转基因轮状病毒疫苗植物表达载体的构建及GUS基因表达

Construction of Rotavirus Vaccine Plant Cell Expression Vector and Expression of GUS Gene

  • 摘要: 目的 选用植物细胞表达轮状病毒结构蛋白,拟构建有效植物细胞表达系统.方法 利用PT-PCR扩增A组轮状病毒外壳蛋白VP4基因片段,经双酶切后与植物表达载体连接,转化感受态细菌TG1.利用地高辛标记探针进行斑点杂交检测,筛选出阳性克隆,拟用该克隆转化马铃薯细胞,研制新型轮状病毒疫苗.本研究还利用β-葡萄糖苷酸酶(GUS)基因转化马铃薯细胞.结果 A组轮状病毒SA11株VP4基因产物大小与设计相同,为960bp.在被检测的未知载体中,有四个显紫色斑,可判定其为带有VP4cDNA的转化载体.用肉眼或显微镜可观察到马铃薯组织细胞中的蓝色物质.结论 在转基因植物组织器官中观察到GUS的活性,获得外源基因转化的条件.拟用本研究构建的表达载体转化马铃薯细胞,研制新型轮状病毒疫苗.

     

    Abstract: Objective Plant cell expression system would be established to express rotavirus structural protein.Methods VP4 gene fragment of rotavirus were amplified by RT-PCR,and were cloned into plant cell expression vector RG-2.The positive clones were selected by dothybridization with Dig-labeled probe.In this study,β-glucuronidase(GUS) were transmitted into potato cells.Results VP4 gene was 960bp as long as designed,and four positive clones were detected and blue matters were found in GUS gene transmitted plant cells.Conclusion GUS activity could be detected in potato cells and a fine transgenic method was established.This gene expression system would be useful for preparing new rotavirus vaccine.

     

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