Tubo R A, Smith H C, Berezney R
Biochim Biophys Acta. 1985 Jul 24;825(3):326-34. doi: 10.1016/0167-4781(85)90020-x.
Alkaline cesium chloride gradient analysis of in vivo [3H]bromodeoxyuridine-labeled and in vitro [alpha-32P]dCTP-labeled DNA was used to determine whether in vitro DNA synthesis in regenerating rat liver nuclei and nuclear matrices continued from sites of replication initiated in vivo. At least 70 and 50% of the products of total nuclear and matrix-bound in vitro DNA synthesis, respectively, were continuations of in vivo initiated replicational forks. The relationship of the in vitro DNA synthetic sites in total nuclei versus the nuclear matrix was examined by using [3H]bromodeoxyuridine triphosphate to density label in vitro synthesized DNA in isolated nuclei and [alpha-32P]dCTP to label DNA synthesized in isolated nuclear matrix. A minimum of about 40% of matrix-bound DNA synthesis continued from sites being used in vitro by isolated nuclei. Furthermore, nuclear matrices prepared from in vitro labeled nuclei were 5-fold enriched in DNA synthesized by the nuclei and were several-fold enriched, compared to total nuclear DNA, in a particularly high density labeled population of DNA molecules.
利用碱性氯化铯梯度分析法对体内[³H]溴脱氧尿苷标记和体外[α-³²P]dCTP标记的DNA进行分析，以确定再生大鼠肝细胞核和核基质中的体外DNA合成是否从体内启动的复制位点继续进行。分别至少70%和50%的总核和与核基质结合的体外DNA合成产物是体内启动的复制叉的延续。通过使用[³H]溴脱氧尿苷三磷酸对分离细胞核中体外合成的DNA进行密度标记，并使用[α-³²P]dCTP对分离核基质中合成的DNA进行标记，研究了总细胞核与核基质中体外DNA合成位点的关系。至少约40%与核基质结合的DNA合成是从分离细胞核在体外使用的位点继续进行的。此外，由体外标记的细胞核制备的核基质中，细胞核合成的DNA富集了5倍，与总核DNA相比，在一个特别高密度标记的DNA分子群体中富集了几倍。
