Lecture 11: Biotechnology 2

Question 1

Biotechnology can improve the human condition through

Answer 1

• treatment of disease
• industrialisation/ resource efficiency e.g. of food processes, clothing & washing industry
• caring for our environment
• improving nutrition

Question 2

Biotchnology can use:

Answer 2

-prokaryotic cells
-single-celled eukaryotes (e.g. yeast)
cell cultures from higher eukaryotes (e.g. Hela cells, plant protoplasts cultures)
-Whole organisms (e.g. transgenic plants & animals)

Question 3

example of biotechnology for treatment of disease

Answer 3

diabetes -INSULIN

Question 4

Insulin:

Answer 4

• first protein to be sequenced
• normally produced in pancreatic beta cells
• Removes excess glucose from the blood
• Injections of insulin can control type 1 diabetes (and also type ll in some cases)
• used to be isolated from bovine and swine pancreas (cattle & pig)

Question 5

synthetic insulin from

Answer 5

the 1960’s

-became the first approved genetically engineered pharmaceutical product in 1982

Question 6

__ ways insulin can be made

Answer 6

two

-expression in E.coli or yeast

Question 7

Method 1 for forming insulin: Chains A & B are produced from two plasmids in two bacterial strains

Answer 7

• Insulin chain A & B
• take purified chains form both (purified chain A and purified chain B)
• place together = disulphide bonds form

Question 8

method 2 for forming insulin : Proinsulin is produced in yeast

Answer 8

proinsulin : A B + C (C joins A + B)

• proinsulin is made and disulfide bonds form
• a protease removes the 33 amino acid chain C, purification

Question 9

DNA sequencing wasn’t invented until the

Answer 9

1970’s

Question 10

PCR wasn’t invented till

Answer 10

1983

Question 11

human genome wasn’t sequenced till

Answer 11

2003

Question 12

Eukaryotic gene can be too ___ to amplify using PCR

Answer 12

LONG
PCR can be used for genes up to about 10 kb
-human genes can range from 200bp - 2,500,000 bp in length

Question 13

genome fragmentation by

Answer 13

restriction enzymes

Question 14

fragments packages into

Answer 14

Bacterial Arificial Chromosomes of 100-300 kb

-amplified bacterial lineages derived from a single bacterium with a single plasmid can be stored

Question 15

how to find your gene:

Answer 15

• Nucleic acid hybridisation can be used to find your gene of interest in a BAC library
• works on the principle of hybridisation of complementary DNA sequences
• radioactive or luminescent probe to identify

Question 16

Industrialisation/ resource efficiency e.g. of food processes, clothing & washing industry how does it help & e.g.

Answer 16

used because it streamlines a process and makes it cheaper and / or more reliable
-Rennet

Question 17

-Cheese production requires the use of

Answer 17

proteases

Question 18

how was proteases traditionally isolated?

Answer 18

as a complex of enzymes from veal calf stomachs

Question 19

1g of extract can coagulate

Answer 19

15litres of milk. Equates to 0.3mg of enzymes per kg cheese

Question 20

how is rennet now sourced?

Answer 20

• chymosin B gene transferred to bacteria & fungi e.g. fungi Aspergilus niger & Kluyveromyces lactis
• Protein produced by fermentation then extracted

Question 21

Advantages to new source of rennet

Answer 21

higher yield
better texture
less bitter

Question 22

Social considerations to new source of rennet:

Answer 22

vegetarian
halla
kosher
(but not organic)

Question 23

plant biotechnology looks at

Answer 23

• increasing productivity
• increasing storage of crop
• increasing nutritional or taste attributes
• disease resistance
• nitrogen fixation for crops
• stress resistance

Question 24

methodology - plant transformation: Naked DNA can be

Answer 24

‘shot’ into cells but vectored DNA is often used to make stable transformations

Question 25

methodology - plant transformation:

Agrobacterium :

Answer 25

transfers part of a plasmid (T-DNA) into plant cells

Question 26

methodology - plant transformation:

Agrobacterium tumefaciens

Answer 26

• plasmid produces plant hormones to induce callus tissue that excretes sugars that the bacteria uses to grow
• disarmed so as not to produce crown galls
• T-DNA can then be used to transfer genes of interest into plant

Question 27

methodology - plant transformation:

Agrobacterium rhizogenes :

Answer 27

-similar infection strategy to A. tumefaciens

• leaf discs or rott sections infected
• hairy roots form
• can induce shoot formation with plant hormones cytokinin and auxin

Question 28

improving nutrition biotech example:

Answer 28

golden rice

Question 29

Golden Rice:

Answer 29

• vitamin D deficiency is a major problem in developing countries
• normal rice ontains no Beta-carotene (provitamin A)

Question 30

to synthesise Beta-carotene in rice grains what needs to be added

Answer 30

2 enzyme

• Phytoene synthase
• Carotene desaturase I

Question 31

phytoene synthase:

Answer 31

• Promoter gt1 (endosperm specific)

- coding region from daffodil

Question 32

Carotene desaturase l:

Answer 32

• promoter gt1 (endosperm specific)

- Coding region from bacterium Pantoea ananatis

Question 33

Golden rice stil needs to be equivalent to normal rice in:

Answer 33

• yield
• disease resistance
• stress resistance
• quality and storage
• available in local varieties (done through introgression)
• insertion in a neutral locus

Question 34

biotechnology can utilise both

Answer 34

prokaryotic and eukaryotic cells and organisms