Nucleotide incorporation across from templating L-nucleoside by DNA polymerases
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- 英文论文题名：Nucleotide incorporation across from templating L-nucleoside by DNA polymerases
- 论文作者：Li Wu ; Yating Xiao ; Qingju Liu ; Yujian He
- 英文论文作者：Li Wu ; Yating Xiao ; Qingju Liu ; Yujian He ; School of Chemistry and Chemical Engineering ; University of Chinese Academy of Sciences
- 年：2016
- 作者机构：School of Chemistry and Chemical Engineering,University of Chinese Academy of Sciences;
- 英文论文关键词：L-thymidine ; Primer extension ; DNA polymerase ; Kinetics ; Mismatch
- 会议召开时间：2016-07-01
- 会议录名称：中国化学会第30届学术年会摘要集-第二十八分会：化学生物学
- 语种：英文
- 分类号：Q75
- 学会代码：ZGHY
- 会议名称：中国化学会第30届学术年会-第二十八分会 ; 化学生物学
- 会议地点：中国辽宁大连
- 主办单位：中国化学会
- 学会名称：中国化学会
- 页数：1
- 文件大小：193k
- 原文格式：D
- 会议级别：全国
摘要
All natural nucleotides and the building blocks of DNA chains possess D-stereochemistry.DNA polymerases are known to select preferably D-nucleotides over L-nucleotides during DNA synthesis.Here,we reported that several DNA polymerases incorporated nucleotides across from L-thymidine in the DNA template.Screening of several DNA polymerases was characterized and the results displayed that primer extension of 5'-3'natural nucleotides gets to end at chiral modification site with Taq and Phanta Max Super-Fidelity DNA polymerase,but 5'-3'primer extension proceeds to the end of the template catalyzed by Deep Vent,Vent and Therminator DNA polymerases.Further,kinetics analyses of primer-extension showed that templating L-nucleosides significantly decreased the nucleotide incorporation rates catalyzed by Deep Vent DNA polymerase or Vent DNA polymerase comparing with normal template,and thus polymerase chain reactions were inhibited with the DNA template containing consecutive L-Ts.In addition,no single base mutation or mismatch mixture corresponding to the location of L-T in the template was found during DNA synthesis when DNA extension pass by L-nucleoside by DNA polymerase.The data reported here offer ample functional grounds to explain why the L-nucleoside might be disfavored as a component of a coding system.
All natural nucleotides and the building blocks of DNA chains possess D-stereochemistry.DNA polymerases are known to select preferably D-nucleotides over L-nucleotides during DNA synthesis.Here,we reported that several DNA polymerases incorporated nucleotides across from L-thymidine in the DNA template.Screening of several DNA polymerases was characterized and the results displayed that primer extension of 5'-3' natural nucleotides gets to end at chiral modification site with Taq and Phanta Max Super-Fidelity DNA polymerase,but 5'-3' primer extension proceeds to the end of the template catalyzed by Deep Vent,Vent and Therminator DNA polymerases.Further,kinetics analyses of primer-extension showed that templating L-nucleosides significantly decreased the nucleotide incorporation rates catalyzed by Deep Vent DNA polymerase or Vent DNA polymerase comparing with normal template,and thus polymerase chain reactions were inhibited with the DNA template containing consecutive L-Ts.In addition,no single base mutation or mismatch mixture corresponding to the location of L-T in the template was found during DNA synthesis when DNA extension pass by L-nucleoside by DNA polymerase.The data reported here offer ample functional grounds to explain why the L-nucleoside might be disfavored as a component of a coding system.
All natural nucleotides and the building blocks of DNA chains possess D-stereochemistry.DNA polymerases are known to select preferably D-nucleotides over L-nucleotides during DNA synthesis.Here,we reported that several DNA polymerases incorporated nucleotides across from L-thymidine in the DNA template.Screening of several DNA polymerases was characterized and the results displayed that primer extension of 5'-3' natural nucleotides gets to end at chiral modification site with Taq and Phanta Max Super-Fidelity DNA polymerase,but 5'-3' primer extension proceeds to the end of the template catalyzed by Deep Vent,Vent and Therminator DNA polymerases.Further,kinetics analyses of primer-extension showed that templating L-nucleosides significantly decreased the nucleotide incorporation rates catalyzed by Deep Vent DNA polymerase or Vent DNA polymerase comparing with normal template,and thus polymerase chain reactions were inhibited with the DNA template containing consecutive L-Ts.In addition,no single base mutation or mismatch mixture corresponding to the location of L-T in the template was found during DNA synthesis when DNA extension pass by L-nucleoside by DNA polymerase.The data reported here offer ample functional grounds to explain why the L-nucleoside might be disfavored as a component of a coding system.
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