title: Using HEK293T Expression System to Study Photoactive Plant Cryptochromes.
abstract: Cryptochromes are photolyase-like blue light receptors that are conserved in plants and animals. Although the light-dependent catalytic mechanism of photolyase is well studied, the photochemical mechanism of cryptochromes remains largely unknown. Lack of an appropriate protein expression system to obtain photochemically active cryptochrome holoproteins is a technical obstacle for the study of plant cryptochromes. We report here an easy-to-use method to express and study Arabidopsis cryptochrome in HEK293T cells. Our results indicate that Arabidopsis cryptochromes expressed in HEK293T are photochemically active. We envision a broad use of this method in the functional investigation of plant proteins, especially in the large-scale analyses of photochemical activities of cryptochromes such as blue light-dependent protein-protein interactions.
authors.affiliation: Laboratory of Soil and Plant Molecular Genetics, College of Plant Science, Jilin UniversityChangchun, China; Basic Forestry and Proteomics Research Center, Haixia Institute of Science and Technology, Fujian Agriculture and Forestry UniversityFuzhou, China.
authors.forename: Liang
authors.initials: L
authors.lastname: Yang
authors.affiliation: Basic Forestry and Proteomics Research Center, Haixia Institute of Science and Technology, Fujian Agriculture and Forestry UniversityFuzhou, China; Department of Molecular, Cell and Developmental Biology, University of California, Los AngelesCA, USA.
authors.forename: Xu
authors.initials: X
authors.lastname: Wang
authors.affiliation: Laboratory of Soil and Plant Molecular Genetics, College of Plant Science, Jilin UniversityChangchun, China; Basic Forestry and Proteomics Research Center, Haixia Institute of Science and Technology, Fujian Agriculture and Forestry UniversityFuzhou, China.
authors.forename: Weixian
authors.initials: W
authors.lastname: Deng
authors.affiliation: Laboratory of Soil and Plant Molecular Genetics, College of Plant Science, Jilin University Changchun, China.
authors.forename: Weiliang
authors.initials: W
authors.lastname: Mo
authors.affiliation: Laboratory of Soil and Plant Molecular Genetics, College of Plant Science, Jilin University Changchun, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Gao
authors.affiliation: Basic Forestry and Proteomics Research Center, Haixia Institute of Science and Technology, Fujian Agriculture and Forestry University Fuzhou, China.
authors.forename: Qing
authors.initials: Q
authors.lastname: Liu
authors.affiliation: Basic Forestry and Proteomics Research Center, Haixia Institute of Science and Technology, Fujian Agriculture and Forestry University Fuzhou, China.
authors.forename: Chuanyu
authors.initials: C
authors.lastname: Zhang
authors.affiliation: Laboratory of Soil and Plant Molecular Genetics, College of Plant Science, Jilin UniversityChangchun, China; Department of Molecular, Cell and Developmental Biology, University of California, Los AngelesCA, USA.
authors.forename: Qin
authors.initials: Q
authors.lastname: Wang
authors.affiliation: Department of Molecular, Cell and Developmental Biology, University of California, Los Angeles CA, USA.
authors.forename: Chentao
authors.initials: C
authors.lastname: Lin
authors.affiliation: Laboratory of Soil and Plant Molecular Genetics, College of Plant Science, Jilin UniversityChangchun, China; Basic Forestry and Proteomics Research Center, Haixia Institute of Science and Technology, Fujian Agriculture and Forestry UniversityFuzhou, China.
authors.forename: Zecheng
authors.initials: Z
authors.lastname: Zuo
keywords: HEK293T; cryptochrome; expression system; linear DNA; photo-biochemical activity