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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 2 Enlarged

Fig. 2:

Map of the bop gene cluster, the minimal bop promoter and promoter mutagenesis. (A) The relative sizes and arrangement of the 4 genes, brp, bat, blp, and bop, in a gene cluster on the chromosome is shown in boxes with position and orientation of the promoters (indicated by arrows above). (B) The sequence of nucleotides within the minimal bop promoter and a few surrounding nucleotides with the TATA box, UAS, and RY box, boxed. The start codon is underlined and the transcription start site (indicated by an arrow) is numbered +1. (C) E. coli-Halobacterium shuttle plasmid (pNB series) with the cloned bop gene (black arrow) and promoter (white box). The plasmid contains origins of replication for E. coli (ori ColE1) and Halobacterium (ori pNRC100, repH) in addition to selectable markers (bla for Ampr in E.coli, and mev for Mevr in Halobacterium [shaded arrows]).

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