Biotechnology (Chapter 20) Objectives

Transcription

1 Biotechnology (Chapter 20) Objectives Understand the background science behind the technology applications Understand the tools and details of the technology Develop familiarity with performing the select techniques

2 Plan Explaining the universality of the genetic material that makes DNA technology possible Brief explanation of use of bacteria in biotechnology applications Technical o Genetic variability in bacteria o Plasmids and plasmid vectors o Restriction Enzymes o Small volumes and proper use of micropipettes o Handling small Eppendorf tubes o Microfuges Overview of Biotechnology

3 DNA Technology & Biotechnology Background Information Definitions of biotechnology, DNA technology, genetic engineering Basic tools of biotechnology: genomic DNA, plasmid vectors, DNA modification enzymes (restriction enzymes) Basic Techniques: DNA gel electrophoresis, transformation, Polymerase Chain Reaction (PCR) Basic Lab Skills (Restriction Enzymes) Plasmid vector components & map Use of micropipette Setting-up restriction enzyme digests of a plasmid

4 Background Information Plasmids Source, composition & architecture Genes found on plasmids Functional DNA sequences of plasmids: o origin of replication o selectable marker- antibiotic resistance gene of interest o Gene(s) to be cloned o Control elements for gene expression Plasmid map Restriction Enzymes Where are they naturally found? Biological function Enzymatic activity- catalysis of sequence-specific cutting of DNA Staggered cutting-sticky ends Blunt cutting-no sticky ends Plasmid Maps with Restriction Enzyme sites Size of plasmid Position of restriction enzymes sites Size of fragments when cut and genes they carry

5 Genetic Material in Bacteria Chromosomal DNA circular Essential genes Extra-chromosomal small circular pieces of DNA- plasmids Non-essential useful genes

6 Plasmids Found in most bacteria, fungi, and plants but not in animal cells Multiply independently of bacterial chromosome Carry useful but non-essential genes, extend the biochemical capabilities of bacteria (antibiotic resistance genes)

7

8 DNA Modification enzymes in Restriction enzymes Endonucleases Bacteria Cleave DNA in a sequence-specific manner Different restriction enzymes cut different sequences Tools for cutting DNA during genetic engineering

9 Definitions Technology: the practical application of science to commerce or industry Biotechnology: the science of using living cells, and components of living cells, to produce goods and services.

10 Name of Bacteria Name of restriction Enzyme Restriction site Escherichia coli EcoR I 5' GAATTC 3' 3' CTTAAG 5' Haemophilus influenzae Serratia marscens Hind III Sma I 5' AAGCTT 3' 3' TTCGAA 5' 5' CCCGGG 3' 3' GGGCCC 5' Restriction enzymes

11 Definitions Engineering: exploiting basic mechanical principles to develop useful tools and objects Genetic engineering: Developing useful tools using genetic material (DNA)

12 Science behind DNA Technology Applications (Lecture topics) Basic understanding of the components of molecular genetics - DNA replication - Transcription - Translation - Control systems regulating all these processes

13 Basic tools of DNA technology - Genomic DNA - Vectors as shuttles for genes between organisms: Plasmids or viruses - Enzymes to manipulate DNA: ligase, DNA polymerase, & Restriction enzymes - Gel electrophoresis

14 Use of plasmids and restriction enzymes in cloning of genes Useful links cloning.html cloning/cloning.htm

15 Biotechnology uses DNA technology to produce useful products

16