Lecture 11: Biotechnology 2 Flashcards

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1
Q

Biotechnology can improve the human condition through

A
  • treatment of disease
  • industrialisation/ resource efficiency e.g. of food processes, clothing & washing industry
  • caring for our environment
  • improving nutrition
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2
Q

Biotchnology can use:

A

-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)

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3
Q

example of biotechnology for treatment of disease

A

diabetes -INSULIN

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4
Q

Insulin:

A
  • 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)
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5
Q

synthetic insulin from

A

the 1960’s

-became the first approved genetically engineered pharmaceutical product in 1982

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6
Q

__ ways insulin can be made

A

two

-expression in E.coli or yeast

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7
Q

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

A
  • Insulin chain A & B
  • take purified chains form both (purified chain A and purified chain B)
  • place together = disulphide bonds form
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8
Q

method 2 for forming insulin : Proinsulin is produced in yeast

A

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
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9
Q

DNA sequencing wasn’t invented until the

A

1970’s

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10
Q

PCR wasn’t invented till

A

1983

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11
Q

human genome wasn’t sequenced till

A

2003

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12
Q

Eukaryotic gene can be too ___ to amplify using PCR

A

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

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13
Q

genome fragmentation by

A

restriction enzymes

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14
Q

fragments packages into

A

Bacterial Arificial Chromosomes of 100-300 kb

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

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15
Q

how to find your gene:

A
  • 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
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16
Q

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

A

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

17
Q

-Cheese production requires the use of

A

proteases

18
Q

how was proteases traditionally isolated?

A

as a complex of enzymes from veal calf stomachs

19
Q

1g of extract can coagulate

A

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

20
Q

how is rennet now sourced?

A
  • chymosin B gene transferred to bacteria & fungi e.g. fungi Aspergilus niger & Kluyveromyces lactis
  • Protein produced by fermentation then extracted
21
Q

Advantages to new source of rennet

A

higher yield
better texture
less bitter

22
Q

Social considerations to new source of rennet:

A

vegetarian
halla
kosher
(but not organic)

23
Q

plant biotechnology looks at

A
  • increasing productivity
  • increasing storage of crop
  • increasing nutritional or taste attributes
  • disease resistance
  • nitrogen fixation for crops
  • stress resistance
24
Q

methodology - plant transformation: Naked DNA can be

A

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

25
Q

methodology - plant transformation:

Agrobacterium :

A

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

26
Q

methodology - plant transformation:

Agrobacterium tumefaciens

A
  • 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
27
Q

methodology - plant transformation:

Agrobacterium rhizogenes :

A

-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
28
Q

improving nutrition biotech example:

A

golden rice

29
Q

Golden Rice:

A
  • vitamin D deficiency is a major problem in developing countries
  • normal rice ontains no Beta-carotene (provitamin A)
30
Q

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

A

2 enzyme

  • Phytoene synthase
  • Carotene desaturase I
31
Q

phytoene synthase:

A
  • Promoter gt1 (endosperm specific)

- coding region from daffodil

32
Q

Carotene desaturase l:

A
  • promoter gt1 (endosperm specific)

- Coding region from bacterium Pantoea ananatis

33
Q

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

A
  • yield
  • disease resistance
  • stress resistance
  • quality and storage
  • available in local varieties (done through introgression)
  • insertion in a neutral locus
34
Q

biotechnology can utilise both

A

prokaryotic and eukaryotic cells and organisms