1. The Structures of DNA and The Structures of DNA and RNARNA

2. GenomeGenomess

3. DNA ReplicationDNA Replication

4. DNA Repair and DNA Repair and RecombinationRecombination

5. Transcription and Transcription and ProcessingProcessing

6. TranslationTranslation

7. Control of Gene ExpressionControl of Gene Expression

8. Manipulating DNA and RNA

Introduction to

Molecular Genetics

http://priede.bf.lu.lv/ Studiju materiāli / MolekularasBiologijas / Ievads MolGen / EN

http://priede.bf.lu.lv/ Studiju materiāli / MolekularasBiologijas / Ievads MolGen / EN

• 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

Some techniques for DNA & RNA analysis

ELECTROPHORESIS separates molecules by size (& structure)

Molecular Biology of the Gene, 5th Edition

HYBRIDIZATION detects specific DNA or RNA sequences

Life The Science of Biology, 7th Edition

The principle of hybridization method

HYBRIDIZATION detects specific DNA or RNA sequences

Localization of genes on chromosomes

HYBRIDIZATION detects specific DNA or RNA sequences

Monitoring the gene expression patterns

GENETIC ENGINEERING combines DNA from different sources

RESTRICTION ENDONUCLEASES cleave specific DNA sequences

GENETIC ENGINEERING combines DNA from different sources

Restriction fragments with compatible sticky ends easily join (anneal)

1

2

3

4

33

Which of the four black sticky ends will anneal to the red sticky end?

GENETIC ENGINEERING combines DNA from different sources

Restriction fragments with compatible sticky ends easily join (anneal)

333

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

GENETIC ENGINEERING combines DNA from different sources

Cloning of a DNA fragment into a plasmid vector

GENETIC ENGINEERING combines DNA from different sources

Amplification of a cloned DNA fragment

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

DNA LIBRARY can be constructed by cloning

There are two types of DNA libraries

Genomic DNA library cDNA library

The synthesis of cDNA

Mutations can be introduced into a gene using MUTAGENESIS

normāls proteīns proteīns, kas satur vēlamo aminoskābes apmaiņu

Genes can be altered in several ways in TRANSGENIC ANIMALS

Genes can be altered in several ways in TRANSGENIC ANIMALS

Genes can be altered in several ways in TRANSGENIC ANIMALS

Reminder on the requirements for DNA synthesis (replication)

Molecular Biology of the Gene, 5th Edition

Polymerase Chain Reaction (PCR) amplifies specific DNA fragments

Molecular Biology of the Cell, 4th Edition

Application of PCR in forensic science

Molecular Biology of the Cell,4th Edition

DNA SEQUENCING determines the nucleotide sequence of DNA fragments

Sanger (1975) dideoxy (chain termination) method

Molecular Biology of the Gene, 5th Edition

DNA SEQUENCING determines the nucleotide sequence of DNA fragments

DNA SEQUENCING determines the nucleotide sequence of DNA fragments

DNA SEQUENCING determines the nucleotide sequence of DNA fragments

Genomes, 2nd Edition

The Human Genome was recently (2001/2004) sequenced