BIO 402502 Advanced Cell amp; Developmental Biology I.ppt

* * * * BIO 402/502 Advanced Cell Developmental Biology I Section IV: Dr. Berezney Lecture 1 The Cell Nucleus and its Genome Organization of Eukaryotic Genome Contrasting features of prokaryotic and eukaryotic genomes with respect to size, percent of coding region and number of genes Renaturation (Hybridization) of DNA DNA renaturation plots for prokaryotic versus eukaryotic DNA demonstrate that: prokaryotic DNA is a unique sequence of DNA whereas eukaryotic DNA is composed of highly repetitive, moderately repetitive and unique sequences. Simple sequence DNA such as satellite DNAs are separated by CsCl density gradients due to major changes in the AT versus CG content (A-T rich DNA has a lower density than GC rich). Alpha Satellite DNA The human alpha satellite sequences at the centromere is an example of tandemly repeated sequences where two chromosomes are held together and connected by spindle fibers for separation of chromosome during mitosis. Gene Structure Introns and Exons : Most of transcribed DNA is intron (~ 90% of the gene sequence), e.g. the chicken ovalbumin gene contains 8 exons 7 introns in over 7.7 kb of DNA. The exons (mRNA) total only 1.9 kb or about 25% of the total transcript, while the factor VIII blood clotting factor gene is 186 kb with 26 exons that compose only about 9 kb or about 5% of the total sequence. Gene Families Pseudogenes Globin gene family; gene amplification: e.g, human type 1 interferon gene cluster is 480 kb in size and is composed of dozens of repeating genes and pseudogenes. Gene duplication or amplification is a result of “unequal crossover” during meiosis is a general mechanism of evolution of tandemly repeated DNA sequences. This is due to misalignment on the two homologous chromosomes. This also leads to gene deletions. Fluorescence In Situ Hybridization (FISH)For detection of specific DNA sequences (e.g., genes) in the nucleus of cells and chromosomes on metaphase spreads  Four step procedu