title: How to Fabricate Functional Artificial Luciferases for Bioassays.
abstract: The present protocol introduces fabrication of artificial luciferases (ALuc(®)) by extracting the consensus amino acids from the alignment of copepod luciferase sequences. The made ALucs have unique sequential identities that are phylogenetically distinctive from those of any existing copepod luciferase. Some ALucs exhibited heat stability, and strong and greatly prolonged optical intensities. The made ALucs are applicable to various bioassays as an optical readout, including live cell imaging, single-chain probes, and bioluminescent tags of antibodies. The present protocol guides on how to fabricate a unique artificial luciferase with designed optical properties and functionalities.
authors.affiliation: Research Institute for Environmental Management Technology, National Institute of Advanced Industrial Science and Technology (AIST), 16-1 Onogawa, Tsukuba, Ibaraki, 305-8569, Japan. kimu-sb@aist.go.jp.
authors.forename: Sung-Bae
authors.initials: SB
authors.lastname: Kim
authors.affiliation: Research Institute for Environmental Management Technology, National Institute of Advanced Industrial Science and Technology (AIST), 16-1 Onogawa, Tsukuba, Ibaraki, 305-8569, Japan.
authors.forename: Rika
authors.initials: R
authors.lastname: Fujii
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001681:Biological Assay; D016466:CHO Cells; D019556:COS Cells; D002522:Chlorocebus aethiops; D003412:Cricetulus; D023061:Gene Order; D008156:Luciferases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D008163:Luminescent Measurements / Q000379:methods*; D010957:Plasmids / Q000235:genetics
chemical_list: D008156:Luciferases
keywords: Artificial luciferase; Bioluminescence; Bioluminescent capsules; Consensus amino acids; Optical readout