Re: Where does the enzyme Pst1 cut the plasmid pBR322?

Hi Susanne -

There are a number of online resources you can use to answer your question.

1. Restriction enzyme cutting sites.

New England Biolabs, among other molecular biology suppliers, includes a listing of restiction enzyme sites in their catalog.

2. Gene sequences for plasmids.

You can query Genbank to obtain the full sequence of pBR322. A query of pBR322 E. coli cloning vector pulled it out, along with sequences from other cloning vectors containing pieces of pBR322.

3. Seek (and destroy)..

With the gene sequence you can search for the nucleotide pattern of the restriction enzyme site. For the quick-and-dirty way, use the search function in the web browser on the gene sequence for pBR322 (Control-F in the PC; Apple-F on the mac). Enter the nucleotide sequence for the enzyme. As most resitrction enzyme sites are palindromic, it doesn't matter wether you're searching the "plus" or "minus" strand of the DNA, if you only have single-stranded DNA. However, if the restriction site crosses a line break, you may miss it with this method. Otherwise, you can scan it manually, or enter it into one of many DNA sequence analysis programs such as DNA Strider which will make the determination for you.

Hope this helps..

-L. Bry, MadSci Admin