The Structures of DNA and RNA Genome s DNA Replication DNA Repair and Recombination

1. Introduction to Molecular Genetics • The Structures of DNA and RNA • Genomes • DNA Replication • DNA Repair and Recombination • Transcription and Processing • Translation • Control of Gene Expression • Manipulating DNA and RNA http://priede.bf.lu.lv/ Studiju materiāli / MolekularasBiologijas / Ievads MolGen / EN

2. Some techniques for DNA & RNA analysis • Electrophoresis • separates molecules by charge, size and structure • Hybridization • detects specific DNA or RNA sequences through complementary base-pairing • Restriction endonucleases • cleave specific double-stranded (ds) DNA sequences • DNA cloning (genetic engineering) • combines DNA from different sources • Reverse transcription • makes DNA (cDNA = copy DNA) from RNA • Mutagenesis • introduces mutations into genes • Transgenic animals • allow elucidation of gene functions • PCR • amplifies specific DNA fragments • Sequencing • determines the nucleotide sequence of nucleic acids

3. ELECTROPHORESIS separates molecules by size (& structure) Molecular Biology of the Gene, 5th Edition

4. HYBRIDIZATION detects specific DNA or RNA sequences The principle of hybridization method Life The Science of Biology, 7th Edition

5. HYBRIDIZATION detects specific DNA or RNA sequences Localization of genes on chromosomes

6. HYBRIDIZATION detects specific DNA or RNA sequences Monitoring the gene expression patterns

7. GENETIC ENGINEERING combines DNA from different sources RESTRICTION ENDONUCLEASES cleave specific DNA sequences

8. GENETIC ENGINEERING combines DNA from different sources Restriction fragments with compatible sticky ends easily join (anneal) 1 2 3 3 4 Which of the four black sticky ends will anneal to the red sticky end?

9. GENETIC ENGINEERING combines DNA from different sources Restriction fragments with compatible sticky ends easily join (anneal) 3 3

10. GENETIC ENGINEERING combines DNA from different sources Vectors transport recombinant DNA into the host cells • A vector should have: • The ability to replicate independently • Restriction sites • Reporter gene(-s) • A relatively small size Life The Science of Biology, 7th Edition

11. GENETIC ENGINEERING combines DNA from different sources Cloning of a DNA fragment into a plasmid vector

12. GENETIC ENGINEERING combines DNA from different sources Amplification of a cloned DNA fragment

13. GENETIC ENGINEERING combines DNA from different sources Expression vectors allow a foreign gene to be expressed in a host cell Life The Science of Biology, 7th Edition

14. DNA LIBRARY can be constructed by cloning

15. There are two types of DNA libraries Genomic DNA library cDNA library

16. The synthesis of cDNA

17. Mutations can be introduced into a gene using MUTAGENESIS normāls proteīns proteīns, kas satur vēlamo aminoskābes apmaiņu

18. Genes can be altered in several ways inTRANSGENIC ANIMALS

19. Genes can be altered in several ways inTRANSGENIC ANIMALS

20. Genes can be altered in several ways inTRANSGENIC ANIMALS

21. Molecular Biology of the Gene, 5th Edition Reminder on the requirements for DNA synthesis (replication)

22. Polymerase Chain Reaction (PCR) amplifies specific DNA fragments Molecular Biology of the Cell, 4th Edition

23. Molecular Biology of the Cell,4th Edition Application of PCR in forensic science

24. DNA SEQUENCING determines the nucleotide sequence of DNA fragments Sanger (1975)dideoxy (chain termination) method Molecular Biology of the Gene, 5th Edition

25. DNA SEQUENCING determines the nucleotide sequence of DNA fragments

26. DNA SEQUENCING determines the nucleotide sequence of DNA fragments

27. DNA SEQUENCING determines the nucleotide sequence of DNA fragments Genomes, 2nd Edition

28. The Human Genome was recently (2001/2004) sequenced