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Cloning and expression of cryptochrome proteins from Arabidopsis thaliana and sunflower (617.3)
Author(s) -
Dyer James,
Li Mengqi
Publication year - 2014
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.28.1_supplement.617.3
Subject(s) - cryptochrome , arabidopsis , arabidopsis thaliana , biology , sunflower , genetics , complementary dna , cloning (programming) , circadian clock , microbiology and biotechnology , gene , mutant , agronomy , computer science , programming language
Two cryptochrome proteins, CRY1 and CRY2, occur in plants and animals. CRY1 regulates the circadian clock in a light‐dependent fashion in insects and plants. Also in plants, blue light photoreception can be used to cue developmental signals. Despite much research on the topic, the specific way that the pterin and flavin chromophores function in these proteins in Arabidopsis thaliana and other plants is still poorly understood. The goal of this research was to clone and express the Arabidopsis and Sunflower CRY1 and CRY2 proteins in an E. coli expression system. Consensus sequence primers flanking the complete coding regions of the two cryptochromes were made based on known sequences of other dicot plant cryptochromes. These primers will be used in a 5’and 3’Rapid Amplification of cDNA Ends protocol, and the complete CRY sequences will cloned into a pBAD‐His vector and purified using Ni‐NTA affinity chromatography. Purified proteins will be used for structural and functional analysis.