从血鞭毛虫原生动物中分离动质体DNA的大环成分。
Isolation of maxicircle component of kinetoplast DNA from hemoflagellate protozoa.
作者信息
Simpson L
出版信息
Proc Natl Acad Sci U S A. 1979 Apr;76(4):1585-8. doi: 10.1073/pnas.76.4.1585.
Maxicircle DNA has been isolated from the kinetoplast DNA of Leishmania tarentolae culture forms by bouyant separation in CsCl in the presence of the A + T-binding dye Hoechst 33258, after liberation from the kinetoplast DNA network by cleavage with the single-hit restriction endonuclease EcoRI. The purified linearized maxicircle DNA has a density in CsCl of 1.681 g/cm3 (79% A + T) and a molecular weight of approximately 18-20 x 10(6). The maxicircle molecule exhibited intramolecular base composition heterogeneity ranging from 85% A + T to 74% A + T.
通过在氯化铯中进行浮力分离，并在存在A+T结合染料Hoechst 33258的情况下，从来自热带利什曼原虫培养形式的动质体DNA中分离出了大环DNA，此前先用单切口限制性内切酶EcoRI切割，使其从动质体DNA网络中释放出来。纯化的线性化大环DNA在氯化铯中的密度为1.681 g/cm³（79%A+T），分子量约为18-20×10⁶。大环分子表现出分子内碱基组成的异质性，范围从85%A+T到74%A+T。
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