通过碱性洗脱法对贻贝血淋巴中DNA交联进行非放射性检测。

Nonradiometric detection of DNA crosslinks in mussel hemolymph by alkaline elution.

作者信息

Batel R, Vukmirović M, Bihari N, Zahn R K, Müller W E

机构信息

Ruder Bosković Institute, Center for Marine Research, Rovinj, Croatia.

出版信息

Anal Biochem. 1993 Aug 1;212(2):402-6. doi: 10.1006/abio.1993.1347.

DOI:10.1006/abio.1993.1347

PMID:7692760

Abstract

The use of bleomycin-Fe(II) complex in the determination of DNA crosslinks by alkaline elution was investigated for the purpose of developing a simple nonradiometric method applicable for detection of DNA damage in marine organisms. The assay involves preparation of active bleomycin-Fe(II) complex and its application in the washing step of the original alkaline elution procedure. Treatment of cell-free DNA from Friend leukemia cells and mussel hemolymph with bleomycin-Fe(II) complex directly on the filter resulted in a concentration-dependent strand scission response with excellent correlation (r = 0.94 and 0.98, respectively). The procedure was validated by determination of DNA cross-links in mussels injected with mitomycine C and formaldehyde.

摘要

为开发一种适用于检测海洋生物DNA损伤的简单非放射性方法，研究了博来霉素-Fe(II)复合物在通过碱性洗脱测定DNA交联中的应用。该测定方法包括制备活性博来霉素-Fe(II)复合物及其在原始碱性洗脱程序的洗涤步骤中的应用。将来自Friend白血病细胞和贻贝血淋巴的无细胞DNA直接在滤膜上用博来霉素-Fe(II)复合物处理，产生了浓度依赖性的链断裂反应，相关性良好（分别为r = 0.94和0.98）。通过测定注射丝裂霉素C和甲醛的贻贝中的DNA交联来验证该程序。