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WHAT IS pBR322?
pBR322 is one of the most used cloning vectors in molecular biology. Cloning vectors, best-known as plasmids, are autonomously replicating DNA units into which DNA fragments can be inserted for gene cloning. Genes taken up by these plasmids are multiplied (or cloned) as the vector replicates, to yields numbers suitable for molecular analysis. The most versatile and well-known plasmid is certainly pBR322 (in fact was one of the first ever used in gene cloning techniques) and has genetically tailored cutting sites into which DNA can be inserted without affecting plasmid self-replication. pBR322 general characteristics are: a) Size: 4.3 kb; b) Replicon: ColE1, relaxed; c) Selective markers (resistance): Amp and Tet; d) Single sites (enzymatic restriction single sites): Ava I, Pst I, BamHI, PvuII, ClaI, SalI, EcoRI, and HindIII.
Explain the functioning of pBR322 vector with diagram?
FUNCTIONG OF pBR322
What is the difference between Puc18 and Puc19?
The multiple cloning sites (MCS) in pUC18 and pUC19 is the difference - the MCS in pUC19 is reverse orientated to those of the pUC18.pUC18: LacZ HindIII PaeI ..... SacI EcoRIpUC19: Lacz EcoRI SacI ..... PaeI HindIII
What is the advantages and disadvantages of pBR322 as a cloning vector?
pBR322 advantages is it widely used for the analysis of prokaryotic transcription and translation as well as topological changes in DNA conformation. then the disadvantage is it has only few cloning sites and the selection procedure is therefore time consuming.
What is the cloning capacity of pBR322 vector?
The cloning capacity of pBR322 vector is 1-5kb.
How many bands were produced after pBR322 was digested with EcoR1?
pBR322 has one EcroR1 site so 1 band however if it was not fully digested you will find 2 or 3 (Linear- [cut], Supercoiled-, Round-Plasmid).
EcoRI and HindIII are What?
They are type II restriction enzymes with a ommon structural core comprising four β-sheets and a single α-helix. restriction endonucleases
What is a degradative enzyme that recognizes specific nucleotide sequences and cuts up DNA?
Those are restriction enzymes, a kind of endonucleases. Some wellknown ones are EcoRI, HindIII or BamHI
How many sites are on the internet?
It is not exactly known how many sites are on the internet,because of personal sites. Well, maybe there are about hundred billions(not sure)
What group of organisms provides a source of restriction enzymes for genetic engineering?
Bacteria. The enzymes are typically named after the bacteria also. For example EcoRI comes from E. coli and HindIII comes from H. influenzae.
Why pBR322 plasmid have high copy number?
I think pBR322 has a replication module from E coli plasmid colE1 ,which permits plasmid replication even when chromosome replication and cell division are inhibited by amino acid starvation and chloramphenicol, as a result, under such condition each cell accumulates several thousands copies of the plasmids up to 3000, so that one litre of bacterial culture easily yields a milligram of plasmid DNA.
How many killing sites were there in the Holocaust?
There were over 100 killing sites
