Fat birds, Fat people, Fat Chance?

http://www.karenfincannon.com/chicbig.jpg

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.

http://www.tulane.edu/%7Edmsander/WWW/335/Adenoviruses.html

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. 2006 Adipogenic potential of multiple human adenoviruses in vivo and in vitro in animals. Jan;290(1):R190-4. Am. J. of Physiol.

• 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. 2005 Jul;43(7):3049-53.

• Database of Ad5 E1A mutations http://www.geocities.com/CapeCanaveral/Hangar/2541/data.html