通过聚（dA）·聚（dT）方法构建的杂交质粒中插入的DNA片段的特异性切除。
Specific excision of the inserted DNA segment from hybrid plasmids constructed by the poly(dA). poly (dT) method.
作者信息
Hofstetter H, Schamböck A, Van Den Berg J, Weissmann C
出版信息
Biochim Biophys Acta. 1976 Dec 13;454(3):587-91. doi: 10.1016/0005-2787(76)90286-0.
The hybrid plasmid PBETAG, consisting of plasmid PMB9 DNA with an insert of rabbit globin DNA (about 600 base pairs) flanked by poly(dA) poly(dT) regions (Maniatis, T., Kee, S.G., Efstratiadis, A. and Kafatos, F.C. (1976) Cell 8, 163-182), was cleaved into two fragments by endonuclease S1 under conditions of partial denaturation. Only the smaller fragment (575 base pairs) contained globin-specific sequences, showing that excision had occurred in the A-T-rich regions. This method of cleavage provides a useful procedure for assessing the length of inserts in hybrid plasmids prepared by the poly(dA)-POLY(DT) tail method, and allows the preparative recovery of the insert.
杂种质粒PBETAG由质粒PMB9 DNA组成，其插入有兔珠蛋白DNA（约600个碱基对），两侧为聚（dA）聚（dT）区域（马尼阿蒂斯，T.，基，S.G.，埃夫斯特拉蒂亚迪斯，A.和卡法托斯，F.C.（1976年）《细胞》8卷，163 - 182页）。在部分变性条件下，该质粒被核酸酶S1切割成两个片段。只有较小的片段（575个碱基对）含有珠蛋白特异性序列，这表明切割发生在富含A - T的区域。这种切割方法为评估通过聚（dA） - 聚（dT）尾法制备的杂种质粒中插入片段的长度提供了一种有用的程序，并且能够对插入片段进行制备性回收。
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