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http://www.mbg.cornell.edu/lis/views/images/rgb.jpg Heat shock proteins HSF Spt6 FISH (Centromere repeat) Indirect immunofluorescence

Heat shock proteins HSF Spt6 - Brandeis Life Sci · Fig. 3. Alignment of the amino acid sequences encoded by the novel Fugu PG1 (fPG1), mousePG1 (musPG1, conceptual translation of

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Page 1: Heat shock proteins HSF Spt6 - Brandeis Life Sci · Fig. 3. Alignment of the amino acid sequences encoded by the novel Fugu PG1 (fPG1), mousePG1 (musPG1, conceptual translation of

http://www.mbg.cornell.edu/lis/views/images/rgb.jpg

Heat shock proteins HSF Spt6

FISH (Centromere repeat)

Indirect immunofluorescence

Page 2: Heat shock proteins HSF Spt6 - Brandeis Life Sci · Fig. 3. Alignment of the amino acid sequences encoded by the novel Fugu PG1 (fPG1), mousePG1 (musPG1, conceptual translation of
Page 3: Heat shock proteins HSF Spt6 - Brandeis Life Sci · Fig. 3. Alignment of the amino acid sequences encoded by the novel Fugu PG1 (fPG1), mousePG1 (musPG1, conceptual translation of
Page 4: Heat shock proteins HSF Spt6 - Brandeis Life Sci · Fig. 3. Alignment of the amino acid sequences encoded by the novel Fugu PG1 (fPG1), mousePG1 (musPG1, conceptual translation of

http://www.sciencemag.org/cgi/content/full/301/5634/793

Page 5: Heat shock proteins HSF Spt6 - Brandeis Life Sci · Fig. 3. Alignment of the amino acid sequences encoded by the novel Fugu PG1 (fPG1), mousePG1 (musPG1, conceptual translation of

http://www.sciencemag.org/cgi/content/full/296/5573/1661

Est = Expression Sequence TagAnchor = homologous segments in unique sequence compared to human

SINES and LINES make up almost half the genome

Page 6: Heat shock proteins HSF Spt6 - Brandeis Life Sci · Fig. 3. Alignment of the amino acid sequences encoded by the novel Fugu PG1 (fPG1), mousePG1 (musPG1, conceptual translation of
Page 7: Heat shock proteins HSF Spt6 - Brandeis Life Sci · Fig. 3. Alignment of the amino acid sequences encoded by the novel Fugu PG1 (fPG1), mousePG1 (musPG1, conceptual translation of
Page 8: Heat shock proteins HSF Spt6 - Brandeis Life Sci · Fig. 3. Alignment of the amino acid sequences encoded by the novel Fugu PG1 (fPG1), mousePG1 (musPG1, conceptual translation of

phage λ gt11 cloning vector: expression of gene fragmentsin E. coli, identified by antibody staining

Page 9: Heat shock proteins HSF Spt6 - Brandeis Life Sci · Fig. 3. Alignment of the amino acid sequences encoded by the novel Fugu PG1 (fPG1), mousePG1 (musPG1, conceptual translation of

pRS316 a yeast/E.coli shuttle vector

http://www.atcc.org/images/vectors/gifs/77145.gif

Page 10: Heat shock proteins HSF Spt6 - Brandeis Life Sci · Fig. 3. Alignment of the amino acid sequences encoded by the novel Fugu PG1 (fPG1), mousePG1 (musPG1, conceptual translation of

F-factor-based BAC cloning vector

Insert here

Page 11: Heat shock proteins HSF Spt6 - Brandeis Life Sci · Fig. 3. Alignment of the amino acid sequences encoded by the novel Fugu PG1 (fPG1), mousePG1 (musPG1, conceptual translation of
Page 12: Heat shock proteins HSF Spt6 - Brandeis Life Sci · Fig. 3. Alignment of the amino acid sequences encoded by the novel Fugu PG1 (fPG1), mousePG1 (musPG1, conceptual translation of
Page 13: Heat shock proteins HSF Spt6 - Brandeis Life Sci · Fig. 3. Alignment of the amino acid sequences encoded by the novel Fugu PG1 (fPG1), mousePG1 (musPG1, conceptual translation of

http://fig.cox.miami.edu/Faculty/Dana/chromosomewalking.jpg

Page 14: Heat shock proteins HSF Spt6 - Brandeis Life Sci · Fig. 3. Alignment of the amino acid sequences encoded by the novel Fugu PG1 (fPG1), mousePG1 (musPG1, conceptual translation of

http://www.entu.cas.cz/fyziol/seminars/genome/walking.jpg

Page 15: Heat shock proteins HSF Spt6 - Brandeis Life Sci · Fig. 3. Alignment of the amino acid sequences encoded by the novel Fugu PG1 (fPG1), mousePG1 (musPG1, conceptual translation of

http://www.fgsc.net/fgn38/cabfig2.gif

Page 16: Heat shock proteins HSF Spt6 - Brandeis Life Sci · Fig. 3. Alignment of the amino acid sequences encoded by the novel Fugu PG1 (fPG1), mousePG1 (musPG1, conceptual translation of
Page 17: Heat shock proteins HSF Spt6 - Brandeis Life Sci · Fig. 3. Alignment of the amino acid sequences encoded by the novel Fugu PG1 (fPG1), mousePG1 (musPG1, conceptual translation of

medlib.med.utah.edu/block2/ biochem/Formosa/menu.html

Page 18: Heat shock proteins HSF Spt6 - Brandeis Life Sci · Fig. 3. Alignment of the amino acid sequences encoded by the novel Fugu PG1 (fPG1), mousePG1 (musPG1, conceptual translation of

RT-PCR AAAAAAAAAAAA mRNA

TTTTTTTTTT

http://www.dur.ac.uk/stefan.przyborski/images/

Page 19: Heat shock proteins HSF Spt6 - Brandeis Life Sci · Fig. 3. Alignment of the amino acid sequences encoded by the novel Fugu PG1 (fPG1), mousePG1 (musPG1, conceptual translation of

http://www.sciencemag.org/cgi/content/full/296/5573/1661

Est = Expression Sequence TagAnchor = homologous segments in unique sequence compared to human

Page 20: Heat shock proteins HSF Spt6 - Brandeis Life Sci · Fig. 3. Alignment of the amino acid sequences encoded by the novel Fugu PG1 (fPG1), mousePG1 (musPG1, conceptual translation of

Ling V, Wu PW, Finnerty HF, Agostino MJ, Graham JR, Chen S, Jussiff JM, Fisk GJ, Miller CP, Collins M.Assembly and annotation of human chromosome 2q33 sequence containing the CD28, CTLA4, and ICOSgene cluster: analysis by computational, comparative, and microarray approaches.Genomics. 2001 Dec;78(3):155-68.

Page 21: Heat shock proteins HSF Spt6 - Brandeis Life Sci · Fig. 3. Alignment of the amino acid sequences encoded by the novel Fugu PG1 (fPG1), mousePG1 (musPG1, conceptual translation of

http://www.sciencemag.org/cgi/content/full/301/5634/793

Where sequences homologous to mouseare found on human chromosomes

what makes the X special?

Page 22: Heat shock proteins HSF Spt6 - Brandeis Life Sci · Fig. 3. Alignment of the amino acid sequences encoded by the novel Fugu PG1 (fPG1), mousePG1 (musPG1, conceptual translation of
Page 23: Heat shock proteins HSF Spt6 - Brandeis Life Sci · Fig. 3. Alignment of the amino acid sequences encoded by the novel Fugu PG1 (fPG1), mousePG1 (musPG1, conceptual translation of

Fig. 3. Alignment of the amino acid sequences encoded by the novel Fugu PG1 (fPG1), mousePG1(musPG1, conceptual translation of mouse cDNA sequence AK003782) and human C9orf21(HumC9orf21) genes. Intron positions (identical in Fugu and humans) are shown by arrowheads. Thehuman C9orf21 sequence has been submitted to GenBank under accession number BK000255.

Page 24: Heat shock proteins HSF Spt6 - Brandeis Life Sci · Fig. 3. Alignment of the amino acid sequences encoded by the novel Fugu PG1 (fPG1), mousePG1 (musPG1, conceptual translation of

evolution of Saccharomyces

S. bayanusS. mikatae

S. paradoxusS. cerevisiae

K. yarowii

S. pombe

20Myr

150Myr

500Myr

Manolis Kellis, Nick Patterson and Chad Nussbaum Whitehead Genome Center

Page 25: Heat shock proteins HSF Spt6 - Brandeis Life Sci · Fig. 3. Alignment of the amino acid sequences encoded by the novel Fugu PG1 (fPG1), mousePG1 (musPG1, conceptual translation of

S.cerevisiae

S.paradoxus

S.mikatae

S.bayanus

Saccharomyces genome alignment

•Local gene order conserved- easy to find orthology•Can study coding and non-coding regions separately

Page 26: Heat shock proteins HSF Spt6 - Brandeis Life Sci · Fig. 3. Alignment of the amino acid sequences encoded by the novel Fugu PG1 (fPG1), mousePG1 (musPG1, conceptual translation of

Where is the genome evolving rapidly?Evolutionary events cluster at

telomeres:

• Rearrangements• Diverged genes

• 80% of changes in 5% ofthe genome

Map rearrangements to cerevisiae genome

Page 27: Heat shock proteins HSF Spt6 - Brandeis Life Sci · Fig. 3. Alignment of the amino acid sequences encoded by the novel Fugu PG1 (fPG1), mousePG1 (musPG1, conceptual translation of

Telomeres: defined regions of rapid change

Spar

Smik

Sbay

Scer

Page 28: Heat shock proteins HSF Spt6 - Brandeis Life Sci · Fig. 3. Alignment of the amino acid sequences encoded by the novel Fugu PG1 (fPG1), mousePG1 (musPG1, conceptual translation of
Page 29: Heat shock proteins HSF Spt6 - Brandeis Life Sci · Fig. 3. Alignment of the amino acid sequences encoded by the novel Fugu PG1 (fPG1), mousePG1 (musPG1, conceptual translation of
Page 30: Heat shock proteins HSF Spt6 - Brandeis Life Sci · Fig. 3. Alignment of the amino acid sequences encoded by the novel Fugu PG1 (fPG1), mousePG1 (musPG1, conceptual translation of