Bio-rad Ligation and Transformation Module Manuel d'utilisateur télécharger pdf (Page 22)

• Its MCS has restriction enzyme sites that can be used for later

manipulation of the DNA

• It is a high copy number plasmid

• It contains the b-lactamase gene,

amp

, which confers resistance to

ampicillin

• It contains the

eco47IR

gene, which allows positive selection of

transformants. This gene codes for the

Eco47I

restriction enzyme, and

when the enzyme is expressed, it is toxic to

E. coli

. When the

eco47IR

gene is disrupted by the insertion of DNA into the cloning site, the gene

will no longer be expressed and the transformed cells will grow and

divide on selective media

• It is 2,974 bp long and its sequence is readily available — the accession

number for pJet1.2 is EF694056

pJet1.2 blunted vector.

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Bio-rad Ligation and Transformation Module Manuel d'utilisateur Page 22