pBR322 and pBR327 Vectors, Essential Properties, Cloning Vectors, Antibiotic Resistance, Plasmid Vectors, Restriction Sites

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Home >> Biotechnology and Genomics >> Cloning and Expression Vectors >>pBR322 and pBR327 Vectors

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pBR322 and pBR327 vectors

The naturally occurring plasmids may not possess all the essential properties of a suitabl cloning vector. Therefore, one may have to restructure them by inserting genes of relaxed replication and genes for antibiotic resistance. This has actually been done and suitable plasmid vectors have been obtained.

One of the standard cloning vectors widely used in gene cloning experiments is pBR322 (derived from E. coli plasmid ColEl), which is 4,362 bp DNA and was derived by several alterations in earlier cloning vectors (pBR322 was named after Bolivar and Rodriguez, who prepared this vector).

It has genes for resistance against two antibiotics (tetracycline and ampicillin), an origin of replication and a variety of restriction sites for cloning of restriction fragments obtained through cleavage with a specific restriction enzyme Another vector pBR327 was derived from pBR322, by deletion of nucleotides between1,427 to 2,516.

These nucleotides were deleted to reduce the size of the vector and to eliminate sequences that were known to interfere with the expression of the cloned DNA in eukaryotic cells pBR327 still contains genes for replication derived from pBR322 and pBR327 used to be very common plasmid vectors but have now been replaced by more efficient and improved vectors.

Plasmid pBR322 and its major characteristic features.

1. Eco RI	2. Sa/I
3. tetracycline resistance	4. origin of replication
5. Pstl	6. amplicillin resistance
7. Pst I	8. EcoRI
9. SalI

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