公开(公告)号：US5503995A

公开(公告)日：1996-04-02

申请号：US261670

申请日：1994-06-17

申请人： Yury Khudyakov ,  Howard A. Fields

发明人： Yury Khudyakov ,  Howard A. Fields

IPC分类号： C12N15/09 ,  C07K14/02 ,  C07K14/18 ,  C12N15/10 ,  C12N15/36 ,  C12Q1/68 ,  C12P19/34

CPC分类号： C12N15/10 ,  C07K14/005 ,  C12Q1/6853 ,  C12N2730/10122 ,  C12N2770/24222

摘要： The invention provides a method for the synthesis of DNA based on a cyclic mechanism of combining deoxyoligonucleotides comprising combining: (a) a series of unique single-stranded deoxypolynucleotides, each having a 5' sequence which, when in double-stranded form, can be enzymatically treated to form a unique 3' single-stranded protrusion for selective cyclic hybridization with another unique single-stranded deoxypolynucleotide of the series; (b) a unique deoxypolynucleotide having a 3' sequence which can selectively hybridize with one of the unique single-stranded deoxypolynucleotides of (a); (c) a polymerase which can direct the formation of double-stranded deoxypolynucleotides from the single-stranded deoxypolynucleotides; and (d) an enzyme which can form a unique single-stranded 3' protrusion from the double-stranded deoxypolynucleotides; under conditions which hybridize the unique single-stranded deoxypolynucleotides in a cyclic manner to form the DNA. Also provided is a kit comprising a series of unique synthesized single-stranded deoxypolynucleotides, each having a 5' sequence which, when in double-stranded form, can be enzymatically treated to form a unique 3' single-stranded protrusion for selective cyclic hybridization with another unique single-stranded deoxypolynucleotide of the series.

摘要翻译： 本发明提供了一种基于组合脱氧寡核苷酸的循环机制合成DNA的方法，包括组合：（a）一系列独特的单链脱氧多核苷酸，每个具有5'序列，当双链形式可以是 酶处理以形成独特的3'单链突起，用于与该系列的另一种独特的单链脱氧多核苷酸进行选择性循环杂交; （b）具有3'序列的独特的脱氧多核苷酸，其可以与（a）的独特的单链脱氧核苷酸之一选择性杂交; （c）可以从单链脱氧寡核苷酸引导形成双链脱氧核苷酸的聚合酶; 和（d）可以从双链脱氧寡核苷酸形成独特的单链3'突起的酶; 在独特的单链脱氧核苷酸以循环方式杂交以形成DNA的条件下。 还提供了一种试剂盒，其包含一系列独特的合成的单链脱氧核苷酸，每个具有5'序列，当以双链形式时，可以进行酶处理以形成用于选择性环状杂交的独特的3'单链突起， 该系列的另一种独特的单链脱氧多核苷酸。