gene basics lecture

29
(Primer on Molecular Genetics. DOE Human Genome 1991-92 Program Report.) The Genome at 4 Levels of Detail

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Molecular biology starts with an understanding of structure and functions of genes and their components,

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Page 1: Gene Basics Lecture

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

The Genome at 4 Levels of Detail

Page 2: Gene Basics Lecture

DNA Contains Four Bases that Base-Pair Complementarily

A-TG-C

Page 3: Gene Basics Lecture

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

Page 4: Gene Basics Lecture

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

The Range of Genome Sizes

Page 5: Gene Basics Lecture

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

DNA Content of Prokaryotes and Eukaryotes

Page 6: Gene Basics Lecture

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

Proportion of SequenceComponents Vary in Eukaryotic Genomes

Page 7: Gene Basics Lecture

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

Page 8: Gene Basics Lecture

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

Many Genes Exist as Families

- Different globin genes are expressed at different developmental stages.

Page 9: Gene Basics Lecture

Nucleus

Plastid

Mitochondria

Compartmentalization of Plant Genetic Material

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

Page 10: Gene Basics Lecture

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

Gene Expression = DNA mRNA Protein

Page 11: Gene Basics Lecture

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

DNA Base Sequence Determines Protein Amino Acid Sequence

Page 12: Gene Basics Lecture

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

Amino Acid Sequence Determines 3-Dimensional Protein Structure

Page 13: Gene Basics Lecture

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

Sickle Cell Anemia and Protein Structure

Page 14: Gene Basics Lecture

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

Genome Sizes, Gene Numbers, and Lethal Loci

Page 15: Gene Basics Lecture

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

Genome Colinearity in Zea mays and

Oryza sativa

Page 16: Gene Basics Lecture

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

Evolutionary Tree Basedon Amino Acid Sequence

for Cytochrome C

Page 17: Gene Basics Lecture

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

Page 18: Gene Basics Lecture

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

Most Eukaryotic Genes Contain Introns (non-coding regions)

Page 19: Gene Basics Lecture

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

Restriction Enzymes

Page 20: Gene Basics Lecture

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

Electrophoresis

Page 21: Gene Basics Lecture

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

DNA Separation byElectrophoresis and

Detection by SouthernHybridization

Page 22: Gene Basics Lecture

Construction of Recombinant DNA Molecules

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

Page 23: Gene Basics Lecture

(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.

Page 24: Gene Basics Lecture

(Alcamo. 1999. DNA Technology,

2nd Ed. Harcourt Press.)

DNA Technology

* Gene Cloning* Protein Expression

Page 25: Gene Basics Lecture

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

PCR:in vitro DNAAmplification

Page 26: Gene Basics Lecture

Construction of Genomicand cDNA Libraries

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

Page 27: Gene Basics Lecture

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

Constructing Clonesfor Sequencing

Page 28: Gene Basics Lecture

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

Cloning a Gene by Chromosome Walking

Page 29: Gene Basics Lecture

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

Types of Genome Maps