摘要
试验旨在探索串联型黄色荧光蛋白(YFP)在真核细胞中的表达,以便于构建用于球虫转基因研究的载体。用PCR方法克隆了不含鼠鸟氨酸脱羧酶PEST序列的目的DNA片段YFP-1和YFP-2,并将其依次连接于原始载体pd2EYFP-N1以替换d2EYFP报告基因,将重组载体电转染Vero细胞,观察荧光情况。结果显示,重组载体pYFP-YFP-N1能够在Vero细胞中正常表达,串联型YFP基因可用于球虫转基因中载体构建。
The experiment was designed to study the expression of tandem yellow fluorescent protein （YFP） in eukaryocyte. DNA fragments of YFP - 1 and YFP - 2 were cloned through ploymerase chain reaction, which did not contain mouse ornithine decarboxylase PEST sequence, and were connected to original vector pd2EYFP -N1 in turn to replace the reporter gene d2EYFP. Subsequently, recombinant vector pYFP -YFP -N1 was transfected into Vero cells by electroporation. The results showed that the recombinant vector pYFP - YFP - N1 was able to express normally in Vero cells, so YFP tandem - type gene can be used to construct vector in transgenesis of coccidian.
出处
《江西农业大学学报》
CAS
CSCD
北大核心
2009年第5期962-966,共5页
Acta Agriculturae Universitatis Jiangxiensis
基金
宁夏大学自然科学研究基金资助项目(ZR0616)
关键词
黄色荧光蛋白
VERO细胞
表达
yellow fluorescent protein（YFP）
Vero cell
expression
