Short communication
نویسندگان
چکیده
We demonstrate that the gel electrophoretic mobility-shift assay (EMSA) can be used for site-selective and quantitative monitoring of nicks in linear double-stranded DNA (dsDNA) thus allowing to expediently follow the nicking activity of enzymes or other agents targeted to a designated dsDNA site. At elevated temperature and/or in the presence of urea, DNA fragments carrying a single nick produced by the nicking enzyme N.BstNBI exhibit a well-detectable gel retardation effect. On the basis of permutation analysis, the decreased electrophoretic mobility of nicked dsDNA fragments is attributed to a bend (or hinge) in the DNA double helix sequence-specifically generated by a nick. Since nick-induced DNA bending depends on interaction between base pairs adjacent to a nick, the change in mobility is different for nicked DNA sites with different sequences. Therefore, EMSA monitoring of differential mobility change caused by nicks within various DNA sequences could be useful for studying the differential base stacking and nearest-neighbor energetics.
منابع مشابه
Short Communication: Haematological and immunological responses of sea bass (Dicentrarchus labrax) to a short-term exposure to increased water levels of nitrate
متن کامل
Short Communication: Haematological and immunological responses of sea bass (Dicentrarchus labrax) to a short-term exposure to increased water levels of nitrate
متن کامل
Short communication: Study on epilithic diatoms in the Balikli Tohma Creek (Darende/Malatya in Turkey)
متن کامل
Short communication: Effects of induced spawning on early development in snout otter clam, Lutraria philippinarum (Deshayes, 1854) (Bivalvia: Mactridae)
متن کامل