LECTURE 14

Question 1

combinant DNA technology steps (5)

Answer 1

1. elucidation of molecular structure of DNA
2. method breaking and joining of DNA
3. identification of a suitable vector or gene carrier
4. availability of a functional host cell for vector
5. effective method of selecting the host cell with rDNA

Question 2

enzyme for cutting DNA?

enzyme for joining them?

Answer 2

1. Restriction Enzyme
2. Ligase

Question 3

common vector used?

Answer 3

plasmid

Question 4

common functional host cell for vector?

Answer 4

animal / plant / bacterial cell

Question 5

4 methods for introducing gene

Answer 5

1. biolistic method
2. electroporation
3. microinjection
4. Agrobacterium tumefaciens

Question 6

rice plants can synthesize ____-_______ but the genes responsible are only active in the ____ and not in the ____

Answer 6

• beta-carotene
• leaves
• grains

*so rDNA turned it on for the rice grains resulting in golden rice

Question 7

what 2 genes were added to make golden rice?

Answer 7

1. phytoene synthase (psy)
2. phytoene desaturase (crt 1)

Question 8

1st gen of GOLDEN RICE

• where did the 2 added genes obtained from

Answer 8

• has psy from daffodils
• crt 1 from Erwinia uredovora

Question 9

2nd gen of GOLDE RICE

• where did the 2 added genes obtained from

Answer 9

• psy from corn and rice

Question 10

tomatoes is high in?

Answer 10

anthocyanin

Question 11

technologies that use living organisms or products from the?

Answer 11

BIOTECHNOLOGY

*began 12000 yrs ago

Question 12

method to produce 2 or more geneticlaly identical organisms?

+ what cells are used?

Answer 12

organismal cloning

• somatic cells

Question 13

first cloned lamb/sheep

Answer 13

dolly

Question 14

production of medically important proteins in the mammary glands of livestock?

Answer 14

molecular pharming

Question 14

targeted mutagenesis?

**introduce mutation

Answer 14

targeted genome editing

(gene disruption/gene insertion of predetermined locus)

Question 15

targeted genome editing use??

Answer 15

site specific nucleases

** Zn Finger Nucleases
** Transcription Activator-like Effector-based Nucleases (TALEN)
**CRISPR-CAS 9

Question 16

short DNA sequence that follows DNA region targeted for cleavage by CRISPR CAS 9?

Answer 16

Protospacer Adjacent Motif

Question 16

APPLICATIONS OF GENETIC ENGINEERING

Answer 16

1. Human Gene Mapping
2. Proteomics
3. Transcriptomics
4. Identification, DNA sequencing, and determination of gene action
5. Human Gene Therapy

Question 16

CRISPR-CAS 9

• 2 key molecules?

Answer 16

1. Single guide RNA
   - consist of crRNA sequence specific to DNA target
   - tracer RNA sequence that interacts with CAS 9 protein
2. Cas9 enzyme
   - endonuclease activity
   - acts as “molecular scissors”

Question 16

CRISPR CAS 9 was used for what disease?

+ what gene was cut off?

Answer 16

THALASSEMIA

• deleted gene BCL11A
  (responsible for suppressing fetal hemoglobin production)

Question 17

2 EXAMPLE OF MISSENSE MUTATION

Answer 17

1. Osteoarthritis (cysteine for arginine)
2. Aortic Aneurysm (arginine for glycine)

*both in alpha 1 collagen gene

Question 18

DISEASE with faulty repair of ds DNA breaks

Answer 18

Breast Cancer Disposition Gene

Question 19

P 53 gene

• functions / effects (3)

Answer 19

1. controls cell cycle (prevents replication of damaged DNA_ )
2. DNA repair
3. apoptosis

Question 20

if there is high amount of DAMAGED DNA, there is high amount of ____ protein

Answer 20

P 53

Question 21

disease with fragile X mental retardation gene?

Answer 21

Fragile X Syndrome (has 200-2000 CGG in mRNA)

Question 22

Human DNA Fingerprinting considers ___ _____ _____

Answer 22

tandemly repeated sequences **each VNTR is given a probability score

Question 23

what hormone was studied in animal genes?

Answer 23

super-ovulation hormone

Question 24

how to map plant genome? + 2 aspects?

Answer 24

use DNA markers 1. exhibits mendelian patterns 2. not subjected to environmental influences

Question 25

EXAMPLES of DNA Markers

Answer 25

1. Restriction Fragment Length Polymorphism (RFLP) 2. Tandem Nucleotide Repeat Markers 3. Polymerase Chain Reaction Marker 4. Random Amplified Polymorphic DNA 5. Single Nucleotide Polymorphism

Question 26

WHAT is "POLYMERASE CHAIN REACTION" ? + 3 steps

Answer 26

in vitro procedure that rapidly and efficiently replicates a fragment DNA from genomic DNA 1. Denaturation 2. Primer annealing 3. Elongation