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Volume 10

Journal of Archives of Medicine

Advanced Biotechnology & Annual Pediatrics 2018

November 28-29, 2018

Novel Trends and Advances in Biotechnology,

Cell & Stem Cell Research

Annual Congress on Pediatrics

World Congress on

November 28-29, 2018 Barcelona, Spain

Joint Event On

Dissecting the regulation of the three tandem promoters of the

Escherichia coli

kps gene cluster at

chromosomal level

Esraa Aldawood and Ian Roberts

University of Manchester, United Kingdom

here are three tandem promoters (PR1-1, PR1-2 and PR1-3) within the PR1

regulatory region of

Escherichia coli

kps capsule gene cluster. In this study, we

dissect out the regulation of the PR1 promoters in the chromosome and examine

the effect of global regulators known to regulate transcription of the kps gene cluster

such as IHF, HNS and slyA. Several constructs were made in UTI89 and UTIP1lacZ

strains with mutations in the -10 of the selected promoter using gene doctoring

followed by K1 phage sensitivity assay and FACS analysis in UTI89 strain and

measuring the transcriptional response using chromosomal lacZ reporter fusion

assay in UIP1lacZ strain.The mutation of PR1-2 UTIP1lacZ reduced β-galactosidase

activity by 50% indicating that PR1-2 is a functional promoter. In contrast mutation of PR1-1 abolished transcription from the

whole PR1 promoter indicating that both PR1-2 and PR1-3 are dependent on PR1-1. These data were mirrored by K1 capsule

production as detected by sensitivity to K1-specific bacteriophage and FACS analysis. Mutation of PR1-2 reduced the capsule

production by half whereas mutation of PR1-1 hindered capsule formation. Moreover, three different mutations in ihf, hns

and slyA were made in those promoter constructs and showed a reduction in the β-galactosidase activity in PR1-2 mutant,

where it has PR1-1 acing, indicating that these regulators may act in PR1-1 specifically. Overall, these results demonstrate the

importance of PR1-1 promoter in the PR1 regulatory region and the complex interplay between multiple promoters. Going

forward I intend to study the individual role of each promoter during infection of particular cell line.

Biography

Esraa Aldawood is pursuing PhD in Molecular Microbiology at the University of Manchester, Uk

Esraa Aldawood et al., Arch Med 2018, Volume 10

DOI: 10.21767/1989-5216-C2-006