Module 1.2 Basic Principles of DNA Recombinant Technology Flashcards
Characteristics of a recombinant DNA molecule
- Formed by laboratory methods of genetic recombination and molecular cloning
- Genetic material from multiple sources
- Creates a DNA molecule that would otherwise not be found in nature
Characteristics of rDNA technology:
- Alteration of genetic material outside an organism and introduction of the altered DNA fragment into organisms via an appropriate vector
- Goal is to obtain enhanced and desired characteristics in living organisms or as their product
- Method of joining two or more DNA molecules o create a hybrid
- the result is a DNA molecule that does not exist in nature
Recombinant DNA technology is made possible by what enzymes?
Restriction endonuclease and ligases
These are enzymes recognizes a specific sequence of DNA and cuts, within, or close to that sequence
restriction endonucleases
Explain the following terms related to restriction endonucleases:
Recognition sequence
Restriction site
Recognition sequence: sequence recognized by restriction endonuclease
Restriction site: location along the DNA where the cut is actually made, may within or outside the recognition site
Which class/es of restriction endonuclease require ATP?
Types I
Which class of restriction endonuclease is most specific? Least specific
Type II (25 bp resolution); Type I (1, 000 bp resolution)
Give examples of restriction endonucleases
EcoR1 (G|AATTC) - from - E. coli R-strain
HindIII (A|AGCTT) - from - Haemophilus influenzae D-strain 3
Joins together fragments of newly synthesized DNA to form a seamless strand
ligase
What are the non-specific counterparts of Restriction Endonucleases?
Deoxyribonucleases and Ribonucleases
This scientist was awarded the 1986 Nobel Prize for medicine in his discoveries on growth factors including nerve growth factors, and epidermal growth factors.
Stanley N. Cohen
What is the first plasmid bacterial cloning vector isolated?
λdvgal 120 (isolated from bacteriophage lambda)
The first cloning vector λdvgal 120 contains genes for what cellular process?
galactose metabolism
λdvgal 120 was concatenated with what genetic material?
Simian Vacuolar Virus 40 (SV40) genome
This new cloning vector was used to create intra- and interspecies rDNA from Salmonella.
plasmid-Stanley Cohen 101 (pSC101),
The first approved gene transfer study was on a child with what disease?
severe combined immunodeficiency disorder