Wikipedia preview

出典(authority):フリー百科事典『ウィキペディア（Wikipedia）』「2015/06/19 18:36:00」(JST)

wiki en

Molecular structure of 2',3-dideoxyadenosine triphosphate (ddATP)

Dideoxynucleotides are chain-elongating inhibitors of DNA polymerase, used in the Sanger method for DNA sequencing. They are also known as 2',3' dideoxynucleotides, and abbreviated as ddNTPs (ddGTP, ddATP, ddTTP and ddCTP).

The absence of the 3'-hydroxyl group means that, after being added by a DNA polymerase to a growing nucleotide chain, no further nucleotides can be added as no phosphodiester bond can be created based on the fact that deoxyribonucleoside triphosphates (which are the building blocks of DNA) allow DNA chain synthesis to occur through a condensation reaction between the 5' phosphate (following the cleavage of pyrophospate) of the current nucleotide with the 3' hydroxyl group of the previous nucleotide. The dideoxyribonucleotides do not have a 3' hydroxyl group, hence no further chain elongation can occur once this dideoxynucleotide is on the chain. This can lead to the termination of the DNA sequence. Thus, these molecules form the basis of the dideoxy chain-termination method of DNA sequencing, which was developed by Frederick Sanger in 1977.^[1]

Dideoxynucleotides are useful in the sequencing of DNA in combination with electrophoresis. A DNA sample that undergoes PCR (polymerase chain reaction) in a mixture containing all four deoxynucleotides and one dideoxynucleotide will produce strands of length equal to the position of each base of the type that complements the type having a dideoxynucleotide present. That is, each nucleotide base of that particular type has a probability of being bonded to not a deoxynucleotide but rather a dideoxynucleotide, which ends chain elongation. Thus, if the sample then undergoes electrophoresis, there will be a band present for each length at which the complement of the dideoxynucleotide is present. It is now common to use fluorescent dideoxynucleotides such that each one of the four has a different fluorescence that can be detected by a sequencer; thus only one reaction is needed.

References

^ Sanger, F., Nicklen, S., and Coulson, A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci, 74(12):5463-7, 1977.

UpToDate Contents

全文を閲覧するには購読必要です。 To read the full text you will need to subscribe.

1. 遺伝学およびゲノミクスのためのツール：細胞遺伝学および分子遺伝学 tools for genetics and genomics cytogenetics and molecular genetics

English Journal

ClickSeq: Fragmentation-Free Next-Generation Sequencing via Click Ligation of Adaptors to Stochastically Terminated 3'-Azido cDNAs.

Routh A1, Head SR2, Ordoukhanian P2, Johnson JE3.
Journal of molecular biology.J Mol Biol.2015 Aug 14;427(16):2610-6. doi: 10.1016/j.jmb.2015.06.011. Epub 2015 Jun 24.
We present a simple method called "ClickSeq" for NGS (next-generation sequencing) library synthesis that uses click chemistry rather than enzymatic reactions for the ligation of Illumina sequencing adaptors. In ClickSeq, randomly primed reverse transcription reactions are supplemented with azido-2',
PMID 26116762

Structural basis of CpG and inhibitory DNA recognition by Toll-like receptor 9.

Ohto U1, Shibata T2, Tanji H1, Ishida H1, Krayukhina E3, Uchiyama S4, Miyake K5, Shimizu T6.
Nature.Nature.2015 Apr 30;520(7549):702-5. doi: 10.1038/nature14138. Epub 2015 Feb 9.
Innate immunity serves as the first line of defence against invading pathogens such as bacteria and viruses. Toll-like receptors (TLRs) are examples of innate immune receptors, which sense specific molecular patterns from pathogens and activate immune responses. TLR9 recognizes bacterial and viral D
PMID 25686612

Studying Bordetella pertussis populations by use of SNPeX, a simple high-throughput single nucleotide polymorphism typing method.

Zeddeman A1, Witteveen S1, Bart MJ1, van Gent M1, van der Heide HG1, Heuvelman KJ1, Schouls LM1, Mooi FR2.
Journal of clinical microbiology.J Clin Microbiol.2015 Mar;53(3):838-46. doi: 10.1128/JCM.02995-14. Epub 2015 Jan 7.
Large outbreaks of pertussis occur despite vaccination. A first step in the analyses of outbreaks is strain typing. However, the typing of Bordetella pertussis, the causative agent of pertussis, is problematic because the available assays are insufficiently discriminatory, not unequivocal, time-cons
PMID 25568442

Japanese Journal

Tailing DNA aptamers with a functional protein by two-step enzymatic reaction(ENZYMOLOGY, PROTEIN ENGINEERING, AND ENZYME TECHNOLOGY)

Journal of bioscience and bioengineering 116(6), 660-665, 2013-12
NAID 110009687827

Hepatic Drug Interaction Between Tacrolimus and Lansoprazole in a Bone Marrow Transplant Patient Receiving Voriconazole and Harboring CYP2C19 and CYP3A5 Heterozygous Mutations