Greenleaf William J, Block Steven M
Department of Applied Physics, Stanford University, Stanford, CA 94305, USA.
Science. 2006 Aug 11;313(5788):801. doi: 10.1126/science.1130105.
We present a method for sequencing DNA that relies on the motion of single RNA polymerase molecules. When a given nucleotide species limits the rate of transcription, polymerase molecules pause at positions corresponding to the rare base. An ultrastable optical trapping apparatus capable of base pair resolution was used to monitor transcription under limiting amounts of each of the four nucleotide species. From the aligned patterns of pauses recorded from as few as four molecules, we determined the DNA sequence. This proof of principle demonstrates that the motion of a processive nucleic acid enzyme may be used to extract sequence information directly from DNA.
我们提出了一种基于单个RNA聚合酶分子运动的DNA测序方法。当特定的核苷酸种类限制转录速率时，聚合酶分子会在与稀有碱基对应的位置暂停。使用一种能够分辨碱基对的超稳定光学捕获装置，在四种核苷酸种类中的每一种处于限量的情况下监测转录过程。从少至四个分子记录的对齐暂停模式中，我们确定了DNA序列。这一原理验证表明，一种进行性核酸酶的运动可用于直接从DNA中提取序列信息。
