I wanted to know if there is any advantage of using pUC 18 and pUC 19 vectors in control of diriection of transcripton since there restriction sites are the same but reversed to each other eventhough transcription is mostly Unidirectional?
Hi there,
pUC plasmids are old and older than PCR. At the time the purpose of 18 and 19 was to give the possibility to clone DNA fragment generated by restriction enzymes in both orientation towards the Plac promoter. Therefore you were sure to have the construct suitable for inducible expression of the ORF potentially present in the fragment.
Hi there,
pUC plasmids are old and older than PCR. At the time the purpose of 18 and 19 was to give the possibility to clone DNA fragment generated by restriction enzymes in both orientation towards the Plac promoter. Therefore you were sure to have the construct suitable for inducible expression of the ORF potentially present in the fragment.
I used the reverse orientation of the MCS to my advantage during 1992 when DNA sequencers were not that easily available. I used these two plasmids to clone my insert in opposite orientation so that using only one primer I couldget a sequence for both the strands.
There are several plasmid-pairs, which are identical in their sequence except a reversed multiple cloning site. As already stated by Dominique and Debdutta, in this way you can chose the orientation of your insert by standard restriction enzyme cloning. Sometimes quite helpful.
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