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The History and Advances of Reversible Terminators Used in New Generations of Sequencing Technology

Genomics, Proteomics & Bioinformatics 2013年第1期11卷 34-40页

作者： Fei Chen Mengxing Dong Meng Ge Lingxiang Zhu Lufeng Ren Guocheng Liu Rong Mu CAS Key Laboratory of Genome Sciences and Information Beijing Institute of GenomicsChinese Academy of Sciences

DNA sequencing using reversible terminators, as one sequencing by synthesis strategy, has garnered a great deal of interest due to its popular application in the second-generation high-throughput DNA sequencing technology. In this review, we provided its history of develop- ment, classification, and working mechanism of this technology. We also outlined the screening strategies for DNA polymerases to accommodate the reversible terminators as substrates during polymerization; particularly, we introduced the ＂REAP＂ method developed by us. At the end of this review, we discussed current limitations of this approach and provided potential solutions to extend its application.

关键词： DNA polymerases Sequencing technology Modified nucleotide primer extension Reversible terminator Sequencing by synthesis

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Analysis of the 5' flanking sequence of the human norepinephrine transporter gene

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Cell Research 1998年第2期8卷 143-149页

作者： HUANG FANG JIAN FEI SHUN KAI MA LI HUAZHU ZHAO PING LIU GUO QIANG CAI ZEN CAN YE LI HE GUO (Shanghai Institute of Cell Biology, Chinese Academy of Sciences, Shanghai 200031, China ) Shanghai Institute of Cell Biology Chinese Academy of Sciences Shanghai China

The human norepinephrine transporter(NET) gene was cloned and structurally analyzed. The far 5' fragment containing exon 1 (a non-coding exon) and exon 2 was sequenced. The transcription start site of the gene in human brain stem tissue was determined by primer extension analysis. It was found that the gene could be transcribed from multiple starting points. The 5' flanking sequence contains a proximal G-C rich region, one possible GSG elemeflt and several SP1 sites. However it does not contain TATA box and CAAT box motifS. Gel shift analysis with nuclear extracts from different tissues of mouse shows that the G-C rich region may be involved in tissue specific expression of the gene.

关键词： Norepinephrine transporter gene primer extension transcription regulation

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Nucleotide incorporation across from templating L-nucleoside by DNA polymerases

Nucleotide incorporation across from templating L-nucleoside...

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中国化学会第30届学术年会-第二十八分会：化学生物学

作者： Li Wu Yating Xiao Qingju Liu Yujian He School of Chemistry and Chemical Engineering University of Chinese Academy of Sciences

All natural nucleotides and the building blocks of DNA chains possess *** polymerases are known to select preferably D-nucleotides over L-nucleotides during DNA ***,we reported that several DNA polymerases incorporated nucleotides across from L-thymidine in the DNA *** 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 ***,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 *** 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 *** data reported here offer ample functional grounds to explain why the L-nucleoside might be disfavored as a component of a coding system.

关键词： L-thymidine primer extension DNA polymerase Kinetics Mismatch

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