Novel green fluorescent protein (GFP) baculovirus expression vectors |
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| Affiliation: | 1. College of Life Sciences, Zhejiang Sci-Tech University, Hangzhou 310018, China;2. Key Laboratory of Applied Marine Biotechnology, Ministry of Education, Ningbo University, Ningbo, 315211, China;3. Key Laboratory of Marine Ecosystem and Biogeochemistry, Second Institute of Oceanography, State Oceanic Administration, Hangzhou 310012, China;1. Department of Agrobiology and Bioresources, Faculty of Agriculture, Utsunomiya University, Mine-machi 350, Utsunomiya-shi, Tochigi 321-8505, Japan;2. Department of Agricultural and Environmental Biology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Yayoi 1-1-1, Bunkyo-ku, Tokyo 113-8657, Japan;3. Genebank, National Institute of Agrobiological Science, Kannondai 2-1-2, Tsukuba-shi, Ibaraki 305-8602, Japan;1. Faculty of Chemistry, University of Wrocław, Joliot-Curie 14, 50-383 Wrocław, Poland;2. Department of Animal Physiology and Development, Institute of Experimental Biology, Adam Mickiewicz University, Umultowska 89, 61-614 Poznań, Poland |
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| Abstract: | Baculovirus expression vectors were constructed, which contained gfp as a reporter gene. Substitutions in the amino acid sequences were carried out to produce two mutant forms of GFP. One of these mutants produces blue color when excited by ultraviolet (UV) light. The other mutant produces a yellow color that can be visualized in regular daylight. The gene of interest cloned in-frame with the gfp open reading frame allows visualization of the produced GFP-fusion protein using UV light. The presence of an in-frame 6 x His tag between the gfp cDNA and the multiple cloning site allows purification of the fusion protein on a Ni-NTA (nickel-nitrilo-triacetic acid) agarose matrix. |
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