Classes of Fatty Acids in Bacteria Class/Example Structure of example 0 l. Saturated: C-(CH2)12-CH3 tetradecanoic acid HO HH II.Unsaturated: C-(CH2)6-C=C-(CH2)6-CH3 omega-7-cis hexadecanoic acid HO 0 CH2 III.Cyclopropane: cis-7,8-methylene C-(CH2)7-C-C-(CH2)5-CH3 hexadecanoic acid HO HH IV.Branched: 13-methyltetradecanoic acid HO -( HCHg 0 H V.Hydroxy: C-CH2-C-(CH2)10-CH3 3-hydroxytetradecanoic acid HO OH (a) 2012 Pearson Education.Inc
FAME Analysis Procedure IDENTIFY ORGANISM ↑ Compare pattern of peaks with patterns in database Bacterial culture Extract fatty acids Peaks from various fatty acid methyl esters Derivatize to form methyl esters Gas chromatography (b) 2012 Pearson Education,inc Each peak from the gas chromatograph is due to one particular fatty acid methyl ester and the peak height is proportional to the amount
FAME Analysis Procedure Each peak from the gas chromatograph is due to one particular fatty acid methyl ester and the peak height is proportional to the amount
16.11 Genotypic Analyses Genomic hybridization as a taxonomic tool基因组杂交法 Organisms to organism 1 Organism 2 be compared: ↓ Genomic DNA Genomic DNA ↓ ↓ DNA preparation Shear and label®) Shear DNA Iee官 Heat to form single ®® strands (a) 2012 Pearson Education,Inc. a)DNA is isolated from test organisms.One of the DNAs is labeled (Organism 1)
Chen Feng, Shanghai Jiao Tong University 16.11 Genotypic Analyses Genomic hybridization as a taxonomic tool 基因组杂交法 a) DNA is isolated from test organisms. One of the DNAs is labeled (Organism 1)
Hybridization experiment: Mix DNA,adding unlabeled DNA in excess: ® 1X1 Hybridized DNA- 1×2 Hybridized DNA- (b) 2012 Pearson Education.Inc. (b)hybridization experiment. Excess unlabeled DNA is added in each experiment to prevent labeled DNA from reannealing with itself.Following hybridization,hybridized DNA is separated from unhybridized DNA before measuring radioactivity in the hybridized DNA only
(b) hybridization experiment. Excess unlabeled DNA is added in each experiment to prevent labeled DNA from reannealing with itself. Following hybridization, hybridized DNA is separated from unhybridized DNA before measuring radioactivity in the hybridized DNA only