Anderson J N, Schimke R T
Cell. 1976 Mar;7(3):331-8. doi: 10.1016/0092-8674(76)90162-8.
Cellulose-bound DNA complementary to ovalbumin mRNA was used in a continous hybridization system to isolate single-stranded DNA molecules containing the ovalbumin gene. Fragmented DNA segments containing the ovalbumin gene were enriched 300-350 fold in one cycle of purification. Two cycles of purification resulted in a DNA fraction which was enriched 2300 fold in the ovalbumin sequence. The method is suitable for purification of the ovalbumin sequence from both sheared DNA fragments, as well as larger molecular weight DNA containing more than twice the number of nucleotides necessary to code for ovalbumin mRNA. The chromatographic procedures were specific and reproducible. In addition, the recovery of ovalbumin DNA was essentially quantitative (80-100%), even when large amounts of starting DNA (70-75 mg) were used. This purification scheme should also be useful for the enrichment of other unique sequence gened form eucaryotic DNA.
与卵清蛋白信使核糖核酸互补的纤维素结合DNA被用于连续杂交系统,以分离含有卵清蛋白基因的单链DNA分子。在一个纯化循环中,含有卵清蛋白基因的DNA片段被富集了300 - 350倍。两个纯化循环产生了一个DNA组分,其在卵清蛋白序列中被富集了2300倍。该方法适用于从剪切的DNA片段以及含有编码卵清蛋白信使核糖核酸所需核苷酸数量两倍以上的更大分子量DNA中纯化卵清蛋白序列。色谱程序具有特异性且可重复。此外,即使使用大量起始DNA(70 - 75毫克),卵清蛋白DNA的回收率基本上是定量的(80 - 100%)。这种纯化方案对于从真核生物DNA中富集其他独特序列基因也应该是有用的。