gene basics lecture

(Primer on Molecular Genetics. DOE Human Genome 1991-92 Program Report.)

The Genome at 4 Levels of Detail

DNA Contains Four Bases that Base-Pair Complementarily

A-TG-C

Species Genome Site (Mbp/1C)

Arabidopsis thaliana 145Oryza sativa ssp. Indica 419-463Sorghum bicolor 748-772Lycopersicon exculentum 907-1000Solanum tuberosum (2n=4X) 1597-1862Nicotiana plumbaginfolia 2287Zea mays 2292-2716Pisum sativum 3947-4397Nicotiana tabacum (2n=4X) 4221-4646Hordeum vulgare 4873Triticum aestivum (2n=6X) 15,966Tulipa sp. 24,704-30,687Homo sapiens 3500Drosophila melanogaster 165Caenorhabditis elegans 100Saccharomyces cerevisiae 4Escherichia coli 4

(Arumuganathan and Earl. 1991. Plant Mol Biol Rep 9:208-218)

Nuclear Genome Sizes

(Cooper. 1997. The Cell. ASM Press.)

The Range of Genome Sizes

(Lewin. 2000. Genes VII. Oxford Universiy Press.)

DNA Content of Prokaryotes and Eukaryotes

(Lewin. 2000. Genes VII. Oxford Universiy Press.)

Proportion of SequenceComponents Vary in Eukaryotic Genomes

AnthocyaninsPhobaphenes

Malonyl-CoA+coumaroyl-CoA

Chalcone

Flavan-4-ol

Flavanone

Dihydroflavonol

A1

Bz1

A1

C2C2

R/B

PI

Vp1

R/B

C1P

Flavan-3,4-diol

3-Hydroxy-anthocyanidine

(ABA)

(Doebley. 1993. Cur Op Gen Devel 3:865-872.)

Genetic Control of Anthocyanin Biosynthesis in Zea mays

(Cooper. 1997. The Cell. ASM Press.)

Many Genes Exist as Families

- Different globin genes are expressed at different developmental stages.

Nucleus

Plastid

Mitochondria

Compartmentalization of Plant Genetic Material

(Westoff et al. 1998. Molecular Plant Development. Oxford University Press.)

(Primer on Molecular Genetics. DOE Human Genome 1991-1992 Program Report.)

Gene Expression = DNA mRNA Protein

(Kreuzer and Massey. 1996. Recombinant DNA Technology. ASM Press.)

DNA Base Sequence Determines Protein Amino Acid Sequence

(Kreuzer and Massey. 1996. Recombinant DNA Technology. ASM Press.)

Amino Acid Sequence Determines 3-Dimensional Protein Structure

(Alcamo. 1999. DNA Technology, 2nd Ed. Harcourt Press.)

Sickle Cell Anemia and Protein Structure

(Lewin. 2000. Genes VII. Oxford Universiy Press.)

Genome Sizes, Gene Numbers, and Lethal Loci

(Ahn and Tanksley. 1993. PNAS 90:7980-7984.)

Genome Colinearity in Zea mays and

Oryza sativa

(Kreuzer and Massey. 1996. Recombinant DNA Technology. ASM Press.)

Evolutionary Tree Basedon Amino Acid Sequence

for Cytochrome C

TRANSCRIPTION

TRANSLATION

Regulatory Elements

Where When HowMuch

Promoter Gene Terminator

STOPGO

mRNA

Protein

Cell Receptor

DNA Encodes Genes and Contains Signals for Their Control

External Signal

(Cooper. 1997. The Cell. ASM Press.)

Most Eukaryotic Genes Contain Introns (non-coding regions)

(Alcamo. 1999. DNA Technology,2nd Ed. Harcourt Press.)

Restriction Enzymes

(Alcamo. 1999. DNA Technology, 2nd Ed. Harcourt Press.)

Electrophoresis

(Alcamo. 1999. DNA Technology,2nd Ed. Harcourt Press.)

DNA Separation byElectrophoresis and

Detection by SouthernHybridization

Construction of Recombinant DNA Molecules

(Alcamo. 1999. DNA Technology, 2nd Ed. Harcourt Press.)

(Alcamo. 1999. DNA Technology, 2nd Ed. Harcourt Press.)

Boyer-Cohen Experimentof 1973

Plasmids are small, circularDNA molecules that can replicate independently in a host cell.

Foreign DNA inserted intoplasmids can generate millions of copies of the inserted gene.

(Alcamo. 1999. DNA Technology,

2nd Ed. Harcourt Press.)

DNA Technology

* Gene Cloning* Protein Expression

(Primer on Molecular Genetics.DOE Human Genome Project1991-1992 Program Report.)

PCR:in vitro DNAAmplification

Construction of Genomicand cDNA Libraries

(Miesfeld. 1999. Applied Molecular Genetics. Wiley-Liss, Inc.)

(Primer on Molecular Genetics.DOE Human Genome Project1991-1992 Program Report.)

Constructing Clonesfor Sequencing

(Primer on Molecular Genetics. DOE Human Genome 1991-1992 Program Report.)

Cloning a Gene by Chromosome Walking

(Primer on Molecular Genetics. DOE Human Genome 1991-1992 Program Report.)

Types of Genome Maps