Biochemical and genetic analysis of nucleic acid-protein interactions

The integration host factor (IHF) of Escherichia coli is a small, sequence-specific DNA-binding protein. The specific and nonspecific DNA binding constants were determined by gel mobility shift assays. The binding constant of IHF for the H$\sp\prime$ site in $\lambda$ attP is 6.8 $\times$ 10$\sp8$ M$\sp{-1}$ and the nonspecific binding constant is 5.8 $\times$ 10$\sp5$ M$\sp{-1}.$ Therefore, the selectivity of IHF binding is about a thousand-fold higher for a specific site over random sequences. To study the molecular determinants specifying IHF binding, a series of 41 oligonucleotides were synthesized containing adenine analogues that modified the surfaces of the major and minor grooves of DNA. The binding constants determined for these analogue-containing H$\sp\prime$ sites revealed that the specific interaction of IHF with its H$\sp\prime$ site involves interactions with both minor and major grooves of the DNA.

Name thesis:degree_name

Ph.D.

Level thesis:degree_level

Dissertation

Discipline thesis:degree_discipline

Microbiology

Grantor

University of Illinois at Urbana-Champaign

Year dc:date

2011

Author dc:creator

• Wang, Siqun

Contributors dc:contributor

• Gardner, Jeffrey F.

• Biology, Molecular
• Biology, Genetics
• Biology, Microbiology

Statement dc:rights

• Copyright 1995 Wang, Siqun

Language dc:language

eng

Identifier

AAI9624530
(UMI)AAI9624530

OAI identifier oai:identifier

oai:www.ideals.illinois.edu:2142/21092