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Author: Baliga, NS
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Methods Article

Promoter analysis by saturation mutagenesis

Nitin S Baliga1*

1 Institute for Systems Biology. 4225 Roosevelt way NE, Suite 200, Seattle, WA 98105. USA. Phone: 206-732-1266 Fax: 206-732-1299

* To whom correspondence should be addressed: Nitin Baliga, Institute for Systems Biology. 4225 Roosevelt way NE, Suite 200, Seattle, WA 98105. USA. Phone: 206-732-1266. Fax: 206-732-1299. Email: nbaliga@systemsbiology.org

Biol. Proced. Online 2001;3:64-69. doi:10.1251/bpo24
Submitted: August 30, 2001; Accepted: December 17, 2001; Published: December 22, 2001.

Indexing terms: promoter regions (genetics); mutagenesis; polymerase chain reaction.

Abbreviations: bop , bacterioopsin.


Figure 3 Enlarged

Fig. 3:

Promoter scanning by saturation mutagenesis. I Tandem oligonucleotides with randomized bases are designed to span the region of interest. II PCR amplification with a primer pair (of which one is mutagenic) yields a collection of copies of the gene downstream to an equally large number of promoter sequences. The number of mutated promoters is equal to the nth exponent of 4 (where n is the number of nucleotides mutated and the randomized base is N = G, A, T, C).

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