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So, it's not unusual for a Nick show to do a run of just 60 episodes.
This data suggested that the inability of Rep5 to function on ITR2 is primarily the sequence of the trs, specifically the requirement for a nick to be generated between G/T.
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Therefore, the strategy for strand discrimination as well as the mechanism for creation of a nick in the daughter strand to initiate excision will be different in case of these organisms.
Overall, the protein responsible for creation of a nick in the absence of MutH to initiate mismatch repair has been identified.
(Thanks to Ben Goldacre. It's pretty much nicked from what he said this morning on Radio 4, but then he does say it all the time, so it's ripe for a nicking).
The requirements are a unique restriction enzyme recognition site which is flanked by sites for a nicking endonuclease (Figure 3).
This problem was eliminated by using a hairpin-containing self-templating oligonucleotide that contains a suicide cassette, a recognition site for a nicking enzyme, and recircularizing and monomerizing the tandem repeated complements by nicking the suicide cassette [7].
The hairpin comprises recognition sequences for a nicking enzyme and a restriction enzyme.
The hairpin is equipped with binding and cleavage sites for a nicking enzyme, which enables nicking and ligation without the addition of a supplementary oligonucleotide.
The circular template is designed with three recognition sites for a nicking enzyme, which cuts the complementary strand with the G-rich tracts.
Besides providing the template for the ligation reaction, the hairpin structure contains two additional features: I) It contains a recognition sequence for a nicking enzyme (a nicking enzyme binds double stranded DNA but cleaves only one strand of the DNA duplex).
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