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A transformation efficiency of 1×10 8 cfu/μg for a small plasmid like pUC19 is roughly equivalent to 1 in 2000 molecules of the plasmid used being introduced into cells. In E. coli , the theoretical limit of transformation efficiency for most commonly used plasmids would be over 1×10 11 cfu/μg.
A pUC19 cloning vector showing the multiple cloning site sequence with restriction enzyme sites. A multiple cloning site (MCS), also called a polylinker, is a short segment of DNA which contains many (up to ~20) restriction sites—a standard feature of engineered plasmids. [1]
(Typical transgene delivery methods involve plasmids, which contain foreign DNA.) The smaller size of minicircles also extends their cloning capacity and facilitates their delivery into cells. Their preparation usually follows a two-step procedure: [4] [5] production of a 'parental plasmid' (bacterial plasmid with eukaryotic inserts) in E. coli
Plasmid sizes vary from 1 to over 1,000 kbp. I doubt that there are any plasmids smaller than roughly 500 bp. Plasmids vary in size; the smallest plasmid is only 846 bp long and contains only one gene. [1] Markus29 13:37, 6 February 2013 (UTC)
They are the standard cloning vectors and the ones most commonly used. Most general plasmids may be used to clone DNA inserts of up to 15 kb in size. One of the earliest commonly used cloning vectors is the pBR322 plasmid. Other cloning vectors include the pUC series of plasmids, and a large number of different cloning plasmid vectors are ...
A conjugative plasmid capable of chromosome integration is also called an episome (a segment of DNA that can exist as a plasmid or become integrated into the chromosome). When conjugation occurs, Hfr cells are very efficient in delivering chromosomal genes of the cell into recipient F − cells, which lack the episome.
For example, pBR322 is a medium copy number plasmid (~20 copies/cell) from which several high copy number cloning vectors (>100 copies/cell) have been derived by mutagenesis, such as the well known pUC series. [1] This delivers the convenience of high plasmid DNA yields but the additional burden of the high copy number restricts the plasmid size.
The bacterial plasmid is a piece of circular DNA which contains regulatory elements allowing for the bacteria to produce a gene product (gene expression) if it is placed in the correct place in the plasmid. The production site is flanked by two restriction enzyme cutting sites "A" and "B" with incompatible sticky ends.