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1. Herbert Boyer discovered that the restriction enzymes of E. coli bacterium can cut the DNA
stands in a particular fashion

2. Restriction enzymes defense mechanism against a bacteriophage by a bacterium
3. They restrict the replication of viral genomes
4. They cut DNA in unique ways called Recognition Sequences or Restriction Sites

5. Stanley Cohen developed a method to remove plasmids from bacterial cells and reinsert them
in other cells
6. Plasmids float freely in cytoplasm and replicate independently from the coding strand of DNA
7. Insertion of recombinant DNA by means of plasmids BIOTECHNOLOGY

8. Creation of recombinant DNA, gene cloning and gene transfer allows the introduction of one or
a set of desired set of genes without introducing undesired genes
9. The alien DNA will link to the origin of replication DNA sequence in the chromosomes and
10. Fragments of DNA are separated through gel electrophoresis
11. Elution extraction of fragments of DNA from the gel piece
12. Selectable marker identify and eliminate non-transformants and selectively permits the
growth of transformants

13. Making the cells competent to take up DNA: Treating cells with divalent cation increases the
efficiency with which DNA enters through the pores of the cell wall
14. Methods of inserting DNA to cells
Incubating cells and DNA on ice Incubating at 42
C - heat shock Incubating on ice
Micro injection DNA directly injected into the animal cells
Gene gun or biolistics cells bombarded with DNA coated micro-particles of Au or W
15. Polymerase Chain Reaction (PCR) produce copies of DNA strands comprises repeated cycles
of heating and cooling

Taq polymerase (blue in colour)
Addition of nucleotides extension of primers
DNA developed from one strand which splits to produce
DNA in cycle 2
16. Bioreactors to process culture
17. Downstream processing separation and purification before marketing
18. Bt Bacillus thuringiensis (bacterium) resistance towards insects
19. RNAi (interference) cellular defense mechanism in eukaryotic organisms
20. RNAi involves silencing an RNA (messenger (m) RNA) using a complementary RNA (small
interference (si) RNA)
21. RNAi involves:
Creation of RISC RNA Induced Silencing Complex
Decodation of cellular mRNA by RISC

22. Oral insulin limitations:
The low pH level (acidic) of stomach breaks down insulin before it reaches the liver pH
level breaks the complex peptide bonds of insulin
Mucus layer of intestine is very thick and passing of insulin through this lining into the
blood is relatively low
23. Applications of biotechnology:
Gene therapy
Molecular diagnosis
Genetically modified crops

24. Sorbent a substance used to sorb (absorb/adsorb) molecules from other substances
25. ELISA Enzyme Linked Immuno-Sorbent Assay
26. GEAC Genetic Engineering Approval/Appraisal Committee

27. Biopiracy - the theft or usurpation of genetic materials especially plants and other biological
materials by the patent process
28. NBA National Biodiversity Authority
Checks biopiracy and protects the indigenous and traditional resources
Application for Intellectual Property Rights related to genetic/biological resources
cannot be made without the approval of NBA
Helps in selection of areas of biodiversity importance by state governments
29. Biodiversity Act 2002 primarily addresses:
Issues of conservation and sustainable use of biological resources in the country
Issues related to genetic resources and associated knowledge
Issues arising from the fair and equitable sharing of benefits from the utilization of
biological resources