What is pUC57 vector?
Description. pUC57 is a common used plasmid cloning vector in E. coli. The vector length is 2,710 bp and is isolated from E. coli strain DH5α by standard procedures.
Is pUC57 high copy?
pUC57 is a derivative of pUC19, and a commonly used cloning vector in E. coli. It has a high copy-number origin, contains an ampicillin resistance cassette, and blue/white colony selection around the multiple cloning site.
How do I know if my plasmid is high or low copy number?
A way to determine experimentally if the copy number of your plasmid is high or low is to perform a miniprep. A high-copy plasmid should yield between 3-5 ug DNA per 1 ml LB culture, while a low-copy plasmid will yield between 0.2-1 ug DNA per ml of LB culture.
What is low copy plasmid?
Many plasmids used for gene cloning and heterologous protein expression in E. coli cells are low copy number or single copy number plasmids. Extraction of these types of plasmids from small bacterial cell cultures produces low DNA yields.
Is pUC18 an expression vector?
Vectors pUC18 and pUC19 are small high-copy number plasmids that are widely used for cloning and manipulation of DNA fragments (9).
What type of vector is pUC18?
pUC18 is a small, high copy cloning vector for replication in E. coli. It has been constructed using the ampicillin resistance gene and the pMB1 origin of replication from pBR322. The pMB1 of pUC18 differs from the pBR322 origin by a single point mutation and the lack of the rop gene, leading to a high copy number.
What do arrows on plasmid mean?
The black arrows show the direction of transcription, which is essential for cloning. If you clone your gene of interest in a middle of another gene, make sure that both of them are transcribed in the same direction. Otherwise, the native promoter can interfere with your gene expression.
What does a plasmid map tell you?
Plasmid maps are graphical representation of plasmids, that show the locations of major identifiable landmarks on DNA like restriction enzyme sites, gene of interest, plasmid name and length etc.
Is there any relation between plasmid size and copy number?
Plasmids vary widely in copy number depending on three main factors: 1) The ori and its constituents – (e.g. ColE1 RNA I and RNA II). 2) The size of the plasmid and its associated insert (bigger inserts and plasmids may be replicated at a lower number as they represent a great metabolic burden for the cell).
Which plasmid has highest copy number?
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.