Module E:  Genomics/Molecular Biology

• Sequencing and cloning DNA
• Genotyping
• Genomics association analyses
• Transfection, infection of cells and tissue
• Real time PCR
• GeneChip (microarray) analysis
• Integration with all other Cores for phenotype analysis

Sequence Comparison Within a Species

Sequencing of Genomic DNA From Three Individuals to Detect Single Nucleotide Polymorphisms (SNPs)

DNA fragments are amplified from genomic DNA using polymerase chain reaction and sequenced using fluorescent dideoxy sequencing on an Applied Biosystems DNA analyzer.

ADRB2 Polymorphisms

Drysdale et al, 2000 PNAS 97(19):10483-10488

Arrangement of DNA polymorphisms (SNPs) in the beta-2-adrenergic receptor gene ADRB2.  SNPs are labeled relative to the first base of the start codon ATG (=1).

Genotopic Association to Asthma Phenotypes in Asthma Case and Control Population (Race Adjusted)

Hawkins et al (2006) Am. J. Resp. Care Crit. Med (Submitted)

Genetic association data showing comparative analysis to SNP genotypes to continuous lung function measurements

                        Response to Therapy Influenced by Genotype

Liggett SB, Assay and Drug Dev. Tech. (2003) 1(2):317-326

Response of individuals to regular treatment with beta-agonist versus as needed treatment, groups (Arg/Arg, Gly/Gly) based on ADBR2 genotype at amino acid 16.  Individuals with the Arg/Arg genotype experienced a decrease in AM peak expiratory flow (PEF) when treated regularly (versus as needed) with beta-agonist, and this decrease is exacerbated upon treatment withdrawal.

                   Raw data from amplification of IFNg RNA using real-time PCR

Use of human, mouse Affymetrix GenChips or custom chips for characterizing gene expression (transcriptome) in cells, tissue.