The construction and contemporary expression of eukaryotic vector of fluorescent protein of G2

• 1、Institute of Bioengineering, Zhejiang Science and Technology University
• 2、Department of Biology and Biochemistry, University of Houston

Abstract：Fluorescent protein is a bioluminescence protein which was discovered orginally from certain coelenterates of the medusa, anthozoa and hydrozoa classes. It is a barrel by elevenβ-sheet formed the wall and the centeralα-helices contained the chromophores and it can give luminescence without any outer facors’assistance except oxygen. Fluorescent protein is stable and can express in heterorganism with little or no cytotoxicity. Besides, it is very easy for it fusing with other proteins for the gene fragement of it is very small (about 700 bp) and it keeps fluorescence after fused with other proteins, so it used expandily as a beacon in the fields of cytology, medical science and molecular biology. Neverthless, fluorecent proteins obtained traditionally restricted in extracting them directly from orgnism or site-directed mutagenesis and random mutation methods and little attentions payed on gene synthesis technology to get the novel fluorescent proteins. Here, we constructed the eukaryotic plasmid of pcDNA3.1(+)-G2-HexaHis based on the novel fluorescent proteins which were synthesized by Microfluidic PicoArray method. The constructed eukaryotic plasmid was then transferred into HeLa cells and the result showed that it can express considerably in the HeLa cells with intensive fluorescence. Besides, the subcelullar location result presented G2 mainly located in plasma and scarcely in nucleolus, so the fluorescent protein of G2 will be a good tool used in localization of proteins in plasma, marking cytoskeleton, cell mitosis and plasma protein expression intensity.

Keywords： eukaryotic plasmid construction transfection subcellular location