Fat birds, Fat people, Fat Chance?

Why are we interested in fat chickens? Study by Whigham et al in Jan 2006

Target audience Non majors for adenovirus problem space Majors or Molecular genetics class: PCR, sequence analysis problem space

Teaching objectives Have students pose a question about adenovirus in humans or animals. Students investigate this question by: 1.Looking at already available sequence data 2.In the lab by collecting specimens from a variety of sources, water, fomites, ocular secretions (depending on your IRB approval)

Some Adenovirus background dsDNA non enveloped icosohedral virus 49 recognized serotypes Variety of diseases in humans infect birds, dogs, cats, goats The genome is not segmented and consists of a single molecule of linear double- stranded DNA.

Some sample questions What is the sequence similarity in human Adenovirus verses ducks, birds, dogs? Is adenovirus correlated with obesity? Does adenovirus exhibit the same diversity as HIV or West Nile viruses? Can adenovirus be amplified from water sources? Farm run off, sewage Can adenovirus be identified from fomites?

What will your students be doing? Reading primary literature Analyzing existing data Bioinformatics, tree builing Collect samples, PCR amplify virus Subclone PCR product. Each student at least one clone Students sequence clone (or send off) Analyze sequences collected within the class and on database.

Ebnar et al 2005 Sample Trees

Techniques used PCR Subcloning Plasmid isolation Gel electrophoresis Sequence analysis Bioinformatics

Timeline Non majors- one lab and follow up A semester long project –Some independent studies students or summer projects to work out the bugs, do the primers work? –Once sequence is collected the sequence data can be entered into a problem space and used in other courses.

PCR primers Many primer sequences directed toward hexon region available –Some accessible online –Or design your own

References Whigham LD, Israel BA, Atkinson RL Adipogenic potential of multiple human adenoviruses in vivo and in vitro in animals. Jan;290(1):R Am. J. of Physiol.Whigham LD, Israel BA, Atkinson RL. Ebner K, Suda M, Watzinger F, Lion T.Related Articles, Molecular detection and quantitative analysis of the entire spectrum of human adenoviruses by a two-reaction real-time PCR assay. J Clin Microbiol Jul;43(7): Ebner K, Suda M, Watzinger F, Lion T.Related Articles, Database of Ad5 E1A mutations r/2541/data.html