Expression, purification of recombinant human cryptochrome I and its application in preparation of protective agent for radiotherapy / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 135-146, 2015.
Article
in Chinese
| WPRIM
| ID: wpr-345519
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.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Plasmids
/
Radiation-Protective Agents
/
Recombinant Proteins
/
Escherichia coli
/
Cryptochromes
Limits:
Humans
Language:
Chinese
Journal:
Chinese Journal of Biotechnology
Year:
2015
Type:
Article
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