摘要
采用gibson组装法构建带有lec-5启动子的lec-5p::gfp::lec-5::lec-53’UTR目的质粒和带有肠道特异性vha-6启动子的vha-6p::gfp::lec-5::lec-53’UTR目的质粒,并通过显微注射方法获得游离型的LEC-5P::GFP::LEC-5荧光蛋白标记虫株和整合型的VHA-6P::GFP::LEC-5荧光蛋白标记虫株。利用激光共聚焦显微镜观察LEC-5在秀丽线虫中的表达和亚细胞定位。结果表明:LEC-5在秀丽线虫的肌肉组织和表皮组织中表达,并位于肠上皮细胞的顶端膜和胞质中的囊泡上。研究结果为进一步深入研究LEC-5在RAB-11介导的上皮细胞极性调控中的作用提供试验依据。
The lec-5p::gfp::lec-5::lec-53'UTR target plasmid with lec-5 promoter and the vha-6p::gfp::lec-5::lec-53'UTR target plasmid with intestinal specific vha-6 promoter were constructed by gibson assembly method.Then,the extrachromosomal strain LEC-5P::GFP:LEC-5 and the integrated strain VHA-6P::GFP::LEC-5 were obtained by microinjection.The expression and subcellular localization of LEC-5 in Caenorhabditis elegans were observed by confocal microscopy.The results showed that LEC-5 was expressed in muscle and epidermal cells of Caenorhabditis elegans and was located at the apical membrane and cytoplasmic vesicles in the intestinal epithelial cells.These results will lay foundation for further study of the role of LEC-5 in RAB-11-mediated epithelial membrane polarity regulation.
作者
张楠
程淑琴
宫鹏涛
李建华
王晓岑
李新
张西臣
ZHANG Nan;CHENG Shuqin;GONG Pengtao;LI Jianhua;WANG Xiaocen;LI Xin;ZHANG Xichen(Key Laboratory for Zoonosis Research of Ministry of Education,Zoonosis Institute of College of Veterinary Medicine,Jilin University,Changchun 130062,China)
出处
《吉林农业大学学报》
CAS
CSCD
北大核心
2024年第2期329-334,共6页
Journal of Jilin Agricultural University
基金
吉林省科技发展计划项目(20190103075JH)。
关键词
秀丽线虫
LEC-5蛋白
GFP标记虫株构建
表达
亚细胞定位
Caenorhabditis elegans
LEC-5 protein
GFP-labeled strain construction
expression
subcellularlocalization