What Particularities Has The Dna Sequence Of The Multicloning Site ?

Question

[Spandankeshav]

Hi all,

Could you please answer this question? I am thinking about it from quite long time but still now I haven't found any satisfied answer and I am devastating by it.

Answer 1

[Pierre Lindenbaum]

For a Cloning vector ? The MCS contains several restriction sites that are recognized by one or more restriction enzymes. Those site are unique in the plasmid , so they are a good place for opening the plasmid and inserting an external DNA fragment there.

[Spandankeshav]

Thanks Pierre, I have found the same answer as you said.

Answer 2

[Larry_Parnell]

This question is difficult to answer given that the question itself is vague and open-ended. There are many examples, mostly word-of-mouth, that the MCS (multi-cloning site) of bacterial (I assume) plasmids can encode transcription terminators or promoters (when placed in the same or other species), cannot always be sequenced in vitro, are suicidal to some bacteria, etc. It is known that one cannot always open up the MCS at adjacent restriction endonuclease sites b/c of physical constraints.

Overall, the thing to keep in mind is this: The MCS is an artificial DNA segment and it can behave - no, receive treatment by its host as such. In other words, keep an eye out for the unexpected; don't be surprised.