Base-Modified Nucleic Acids as a Powerful Tool for Synthetic Biology and Biotechnology.
作者： Elena Eremeeva , Michail Abramov , Lia Margamuljana , Piet Herdewijn
关键词: Chemistry 、 Nucleoside triphosphate 、 Nucleic acid analogue 、 Biochemistry 、 Real-time polymerase chain reaction 、 DNA 、 Polymerase cycling assembly 、 Restriction enzyme 、 Oligonucleotide 、 Endonuclease
摘要: The ability of various nucleoside triphosphate analogues deoxyguanosine and deoxycytidine with 7-deazadeoxyadenosine (A1 ) 5-chlorodeoxyuridine (T1 to serve as substrates for Taq DNA polymerase was evaluated. set composed A1 , T1 7-deazadeoxyguanosine either 5-methyldeoxycytidine or 5-fluorodeoxycytidine successfully employed in the chain reaction (PCR) 1.5 kb fragments well random oligonucleotide libraries. Another effective combination triphosphates synthesis a 1 PCR product deoxyinosine, 5-bromodeoxycytidine. In vivo experiments using an antibiotic-resistant gene containing latter demonstrated that bacterial machinery accepts fully modified sequences genetic templates. Moreover, base-modified segments selectively protect from cleavage by restriction endonucleases shown. This approach can be used regulate endonuclease pattern.
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