title: [Expression, purification of recombinant human cryptochrome I and its application in preparation of protective agent for radiotherapy].
abstract: Radiotherapy is a treatment for cancer with undesired by-effects. In order to develop a new radiation protective agent that could reduce the by-effects, we tried to express and purify human cryptochrome 1 (hCRY1). The coding sequence of hCRY1 was inserted into prokaryotic expression plasmid pET28a(+), and this protein was purified from Escherichia coli BL21(DE3) after IPTG induction, ultrasonication, inclusion body dissolution, gradient dialysis, nickel column purification and ultrafiltration. The yield of hCRY1 in 1 L E. coli culture (LB medium) was about 10-15 mg. The radiation protective efficiency of hCRY1 was monitored by detecting X-ray-induced H2A.X foci in HaCaT cells. The results of immunofluorescence show that hCRY1 significantly reduces X-ray stimulated DNA damage response. The apoptosis of HaCaT cell was also detected, and the repression of H2A.X foci formation was not due to hCRY1's cytotoxity. All these data suggest a potential application of recombinant hCRY1 as a protective agent for radiotherapy.
authors.forename: Chen
authors.initials: C
authors.lastname: Yao
authors.forename: Chunjie
authors.initials: C
authors.lastname: Sheng
authors.forename: Dong
authors.initials: D
authors.lastname: Liu
authors.forename: Shijuan
authors.initials: S
authors.lastname: Gao
authors.forename: Wei
authors.initials: W
authors.lastname: Jiang
authors.forename: Hongyan
authors.initials: H
authors.lastname: Yu
authors.forename: Jiandong
authors.initials: J
authors.lastname: Li
authors.forename: Huiming
authors.initials: H
authors.lastname: Chen
authors.forename: Jiaoxiang
authors.initials: J
authors.lastname: Wu
authors.forename: Changchuan
authors.initials: C
authors.lastname: Pan
authors.forename: Shuai
authors.initials: S
authors.lastname: Chen
authors.forename: Wenlin
authors.initials: W
authors.lastname: Huang
mesh_terms: D056931:Cryptochromes / Q000096:biosynthesis*; D004926:Escherichia coli; D006801:Humans; D010957:Plasmids; D011837:Radiation-Protective Agents; D011994:Recombinant Proteins / Q000096:biosynthesis*
chemical_list: C538963:CRY1 protein, human; D056931:Cryptochromes; D011837:Radiation-Protective Agents; D011994:Recombinant Proteins