Horton R M, Cai Z L, Ho S N, Pease L R
Dept. of Biochemistry and Molecular Biology, Mayo Clinic, Rochester, MN 55905.
Biotechniques. 1990 May;8(5):528-35.
Gene Splicing by Overlap Extension or "gene SOEing" is a PCR-based method of recombining DNA sequences without reliance on restriction sites and of directly generating mutated DNA fragments in vitro. By modifying the sequences incorporated into the 5'-ends of the primers, any pair of polymerase chain reaction products can be made to share a common sequence at one end. Under polymerase chain reaction conditions, the common sequence allows strands from two different fragments to hybridize to one another, forming an overlap. Extension of this overlap by DNA polymerase yields a recombinant molecule. This powerful and technically simple approach offers many advantages over conventional approaches for manipulating gene sequences.
重叠延伸基因剪接或“基因SOEing”是一种基于聚合酶链式反应（PCR）的方法，用于在不依赖限制性酶切位点的情况下重组DNA序列，并直接在体外产生突变的DNA片段。通过修饰掺入引物5'端的序列，任何一对聚合酶链式反应产物都可以在一端共享一个共同序列。在聚合酶链式反应条件下，共同序列允许来自两个不同片段的链相互杂交，形成重叠。DNA聚合酶对这种重叠的延伸产生一个重组分子。与传统的基因序列操作方法相比，这种强大且技术简单的方法具有许多优势。
