Week 1-3rd Qtr Genetic Engineering PDF

GENERAL BIOLOGY 2 Genetic engineering is the artificial manipulation, modification, and recombination of DNA or other nucleic acid molecules in order to modify an organism or population of organisms. As such, the term embraced both artificial selection and all the interventions of biomedic...

GENERAL BIOLOGY 2 Genetic engineering is the artificial manipulation, modification, and recombination of DNA or other nucleic acid molecules in order to modify an organism or population of organisms. As such, the term embraced both artificial selection and all the interventions of biomedical techniques, among them artificial insemination, in vitro fertilization, cloning, and gene manipulation.  Genetic engineering is the process of using recombinant DNA (rDNA) technology to alter the genetic makeup of an organism. Traditionally, humans have manipulated genomes indirectly by controlling breeding and selecting offspring with desired traits. Genetic engineering involves the direct manipulation of one or more genes. Classical plant breeding uses deliberate interbreeding (crossing) of closely or distantly related individuals to produce new crop varieties or lines with desirable properties. Genetic engineering involves the use of molecular techniques to modify the traits of a target organism. The modification of traits may involve: 1. introduction of new traits into an organism 2. enhancement of a present trait by increasing the expression of the desired gene 3. enhancement of a present trait by disrupting the inhibition of the desired genes’ expression. The three modes of gene transfer between bacteria are TRANSDUCTION, which is transfer via virus; TRANSFORMATION, which is the uptake of DNA from the surrounding fluid and CONJUGATION, which is the bacterial version of mating.  General Outline of Recombinant DNA 2. selection of an appropriate vector 3. ligation (join or vehicle which together) of the 1. cutting or would propagate gene of interest cleavage of the recombinant (eg. from animal) DNA by DNA (eg. circular with the vector (cut restriction plasmid in bacteria bacterial with a foreign gene plasmid) enzymes of interest) (REs) 1. cutting or cleavage of DNA by restriction enzymes (REs)  General Outline of Recombinant DNA 5. selection 6. sequencing 4. transfer of the process to of the gene to recombinant screen which find out the plasmid into a cells actually primary host cell (that contain the structure of would carry out gene of the protein replication to interest make huge copies of the recombined plasmid) Ways in which these plasmids may be introduced into host organisms: Biolistics In this technique, a “gene gun” is used to fire DNA-coated pellets on plant tissues. Plasmid insertion by Heat Shock Treatment Heat Shock Treatment is a process used to transfer plasmid DNA into bacteria. Cells are incubated with the desired plasmid for about 30min Electroporation This technique follows a similar methodology as Heat Shock Treatment, but, the expansion of the membrane pores is done through an electric “shock”. Some methods are: Selection of plasmid DNA containing cells Selection of transformed cells with the desired gene PCR detection of plasmid DNA Genetically Modified Organisms (GMOs)