RNA polymerase transcription promoter and nucleic acid sequencing method

Abstract

An RNA polymerase transcription accelerator comprising a compound represented by the following Formula (I) or salts thereof. A method of sequencing DNA in which nucleic acid transcripts are obtained using an RNA polymerase and a DNA fragment as a template, the resulted nucleic acid transcripts are separated, the nucleic acid sequence is determined from the separated fractions wherein the nucleic acid transcription reaction is carried out in the presence of a compound selected from a group of compounds represented by the above formula (I). The polyamine compounds above have outstanding accelerating activity on transcription activity of RNA polymerase. Therefore, use of the polyamine compounds in a DNA sequencing method using RNA polymerase can make a length of DNA sequence that can be determined in one sequencing longer.

1 . An RNA polymerase transcription accelerator comprising a compound represented by the following Formula (I) or salts thereof.  
       
         
           
           
               
               
           
         
       
       wherein: 
 n is an integer of 1-8,  
 R 1  represents a hydrogen atom or a p-toluenesulfonyl group,  
 R 2  represents an ethyl group or a group represented by the following Formula (II),  
 R 3  represents a hydrogen atom or a p-toluenesulfonyl group, and  
 R 4  represents an ethyl group or a group represented by the following Formula (II):  
                     
 wherein, m is 1 or 2,  
 R 5  represents a hydrogen atom, and  
 R 6  represents a hydrogen atom or an ethyl group.  
 
     
     
         2 . A method for sequencing DNA in which nucleic acid transcripts are produced by using RNA polymerase and a DNA fragment as a template, the resulting nucleic acid transcripts are separated, and the nucleic acid sequence is determined from the separated fractions, characterized in that said nucleic acid transcription reaction is carried out in the presence of at least one compound selected from the group of compounds according to  claim 1 .  
     
     
         3 . The method of  claim 2  wherein said DNA fragment comprises a promoter sequence for the RNA polymerase, and the nucleic acid transcription reaction is carried out using ribonucleoside-5′-triphosphates comprising ATP, GTP, CTP and UTP or derivatives thereof, and one or more kinds of 3′-deoxyribonucleoside-5′-triphosphate (hereinafter referred to as 3′ dNTP derivatives) comprising 3′ dATP, 3′ dGTP, 3′ dCTP, 3′ dUTP and derivatives thereof.  
     
     
         4 . The method of  claim 2  wherein the 3′ dNTP derivatives are labeled, and nucleic acid sequence is determined using the label.  
     
     
         5 . The method of  claim 3  wherein the 3′ dNTP derivatives are labeled, and nucleic acid sequence is determined using the label.