TAILIEUCHUNG - Báo cáo khoa học: Determination of the reopening temperature of a DNA hairpin structure in vitro

A novel method, based upon primer extension, has been developed for measuring the reopening temperature of a single type of DNA hairpin structure. Two DNA oligo-nucleotides have been utilized and designated as primers 1 and3¢-termini fully comple-mentary to the hairpin flanking sequences, was used to evaluate primer extension conditions, and primer 2, with its 3¢-end competing with the DNA hairpin stem, was used to detect the DNA hairpin reopening temperature. | Eur. J. Biochem. 271 3665-3670 2004 FEBS 2004 doi Determination of the reopening temperature of a DNA hairpin structure in vitro Xuefeng Pan Institute of Microbiology The Chinese Academy of Sciences Beijing China A novel method based upon primer extension has been developed for measuring the reopening temperature of a single type of DNA hairpin structure. Two DNA oligonucleotides have been utilized and designated as primers 1 and 2. Primer 1 with its 5- and 3 -termini fully complementary to the hairpin flanking sequences was used to evaluate primer extension conditions and primer 2 with its 3 -end competing with the DNA hairpin stem was used to detect the DNA hairpin reopening temperature. A single DNA hairpin structure was formed on the DNA template by thermal denaturation and renaturation and this hairpin structure was predicted to prevent the annealing of the 3 -end of primer 2 with the template DNA which leads to no primer extension. By incubating at different temperatures the DNA hairpin structure can be reopened at a particular temperature where the primer extension can be carried out. This resulted in the appearance of double-stranded DNA that was detected on an agarose gel. This temperature is defined here as the hairpin reopening temperature. Keywords DNA hairpin non-B DNA secondary structure primer extension reopening temperature Tm. The significance of DNA folding into non-B secondary structures . pseudohairpin hairpin palindromic triplex and G-tertraplex DNA molecules is twofold. Firstly DNA non-B secondary structures play important physiological roles. For example some GC-rich DNA sequences that fold into G-tertraplex structures participate in the regulation of gene expression 1 and the maintenance of telomere structures 2-4 and some AT-rich DNA sequences in the DNA replication origins of bacterial plasmids that can adopt non-H bounded conformations control DNA replication initiation 5 . Moreover certain types of .

• Báo cáo khoa học
• Report medicine
• traditional medicine
• scientific reports
• genetics
• Structural biochemistry
• Structural Biology
• medical knowledge
• medical research
• analysis of cytoplasmic
• oxidation
• Crystal structure
• bacteria
• hemoglobin
• medicine
• cell proliferation
• breast cancer
• chemical reaction
• gastric cancer
• hormone medicine
• tumor cells
• biochemical studies
• environmental medicine