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Biochemistry > M2: Molecular Genetics & Biotechnology > Flashcards

M2: Molecular Genetics & Biotechnology Flashcards

1
Q

What is a gene?

A

Genes are made up of DNA and are inherited by parents

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2
Q

What is the “central dogma of molecular biology”?

A 
The flow of genetic information in the cell. 
> Transcription control 
> RNA processing and stability 
> Tranlational control 
> Protein processing 
> Protein activity and stability
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3
Q

What is transcriptional control and why is it important?

A

Determines when and in what cells a gene is transcribed to produce mRNA. It is is the 1st step determining how many individual proteins are produced in a cell

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4
Q

If every cell in the body has the same genetic code, how is it that different organs of the body are so unique?

A

about 1000-2000 genes are unique to a specific cell type

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5
Q

Briefly outline the process of transcription

A

The process where a DNA sequence is copied into an RNA molecule

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6
Q

Briefly outline the process of translation

A

synthesis of proteins

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7
Q

What are the key regulatory elements of a gene?

A

> Promoter - DNA sequence at which transcription factors bind and recruit RNA polymerase
AUG
UAG

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8
Q

What are the key elements of the transcribed region of a gene?

A

Transcribed region - sequences of DNA that are copied into RNA.
The transcribed RNA is processed so that it can be translated

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9
Q

What is a transcription factor and how do they control gene expression?

A

Gene is only expressed when both activator and transcription factors are present and the repressor is absent

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10
Q

what are Mendel’s law of inheritance?

A
  1. Law of Segregation
  2. Law of independent assortment
  3. Law of dominance
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11
Q

What is the difference between heterozygous and homozygous mutation, and how would each be inherited?

A

Heterozygous:
> Only one of the enzymes have been knocked out (from either mum or dad) and the other copy has sufficient enzyme to catalyse the reaction.
Homozygous:
> Both enzymes form mum and dad have been knocked out. So the enzyme before will do their job up until here and form a cumulation. The enzymes after will not have any substrate to work with so will have no effect.

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12
Q

From a genetic perspective, why are we all different?

A

Unique combination of alleles from out parents

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13
Q

PKU is a disorder of phenylalanine breakdown. How does altering a single protein in the phenylalanine breakdown pathway lead to disease?

A

It causes the amino acid phenylalanine to build up in the body

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14
Q

What are the major symptoms of PKU and how does this relate to the defect in the processing pathway?

A

black urine - homozygous mutations of the enzyme leading to the pathway not being completed (?)

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15
Q

With regards to to mutations in enzymes involved in metabolic pathways, would a mutation to an enzyme near the start of the pathway, or near the bottom be expected to lead to a more severe phenotype?

A

end (?)

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16
Q

What is the link between genotype and phenotype?

A

Genotype determines pehnotype

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17
Q

How can phenotype be altered by the environment?

A

providing too much or not enough of something

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18
Q

Briefly outline the process of translation

A

A genes is transcribes and mRNA is produced. mRNA goes to the cytoplasms where the ribosomes are . The ribosomes bind to the end of mRNA and scans along until it find the start (AUG) codon. Base pairing between tRNA and codon. mRNA is translated through the actions of tRNA recognising the codons. Release factor is a protein and is able to recognise the stop codon.

19
Q

Use the codon table to determine the amino acids encoded by the DNA sequence CAC and GTA

A

CAC -> GTG (Val)

GTA -> CAT (His)

20
Q

How could a genetic test for a specific disease-causing mutation . be carried using PCCR?

A

Heating to allow for the polymerase to be added and then cooled to allow the primers to bind. This happens on re[peat to form many copies.

21
Q

Why is it important to tightly control cell number?

A

could lead to cancer

22
Q

What is the difference between germline mutations and somatic mutations?

A

Germline mutations are found in gamete cells and can be passed onto the next generation.
Somatic Mutation occur in other cells and cannot be passed on

23
Q

What is cancer?

A

collection of related diseases. Cancer cells are cells within the tissue that longer respond to many of the signals. Over time these can become increasingly resistant. In the late stages, it can break through normal tissue boundaries and spread to other regions

24
Q

What 2 types of genes cause cancer when mutated?

A

Tumor suppressor genes

Onco-genes

25
Q

What is the normal function of these 2 genes?

A

Tumor suppressor genes:
> encode proteins that normally prevent uncontrolled cell growth
> recessive

Onco-genes
> Encode proteins that promote cell growth
> dominant

26
Q

Regarding these classes of genes, under what circumstance would mutations result in cancer?

A

Tumor suppressor genes:
> lack of these genes can stimulate cell growth and is recessive mutation promoting cancer

Onco-genes:
> dominant mutation promoting cancer

27
Q

Briefly discuss the common mutations that lead to chronic myeloid leukaemia, including how these mutations lead to uncontrolled cell divisions

A

inherited mutations increase chance of cancer
> Inherited (germline) mutation a tumor suppressors gene
> Mutation in second copy of tumor suppressor gene

28
Q

In general, how do cancer treatments work?

A

Kill the excess cells

29
Q

What are recombinant DNA technologies?

A

Joining different DNA fragments together and then inserting them into host organism that will produce your protein of interest.

30
Q

What is the crucial element in recombinant DNA technologies?

A

Plasmids

31
Q

What are the key components of plasmids?

A

> circular pieces of dsDNA
replicates independently of the hosts chromosomal DNA
provides benefit of hosts

32
Q

What are restrictive enzymes?

A

defense system to degrade foreign DNA

33
Q

What are DNA ligases?

A

catalyse the formation of phosphodiester bond to repair nick in DNA backbone

34
Q

How are restriction enzymes and DNA ligases useful in recombinant DNA technologies?

A

allows for amplification

35
Q

Briefly outline the process of transformation

A

“transfer of plasmids into bacteria”
> transformation of plasmid to bacterial cell
> plasmid in cell expresses antibiotic resistance gene
> PLate cells on LB Agar antibiotic plate
> Only the cells that contain the plasmid will be able to grow/divide and form colonies

36
Q

How is recombinant DNA technology made possible via the genetic code?

A

we can transform a human gene into bacteria and it will make the same protein

37
Q

What issue may be encountered when cloning eukaryotic genes in prokaryotes and how is this issue overcome?

A

prokaryotic genes do not have introns –> therefore only use the coding sequence

38
Q

What are the steps involved in producing a recombinant protein?

A
  1. Isolate gene of interest
  2. Clone into expression plasmid - which one?
  3. Transform into bacteria for expression or isolation of more DNA for use in another expression system
  4. Grow cells expressing protein of interest
  5. Isolate and purify the protein
39
Q

What are the advantages and disadvantages of using a prokaryotic system to generate human recombinant proteins?

A 
Advantages:
> relatively low cost 
> High yield 
> Pathogen free
Disadvantages:
> Proteins often partially folded
> inability to preform post-translational modification
40
Q

What advantages does a whole animal system have over a cell culture system in the generation of recombinant proteins?

A

cells in culture cannot preform all post-translation modification equally well

41
Q

What factors would you take into consideration when choosing a system to produce your recombinant protein of interest?

A

financial and biochemical reasons

42
Q

In general terms, how do we engineer proteins for biotechnology?

A

Optogenetics
> cell type specific promoter
> Chamelerhedopsin activates
> Antibiotic resistance gene (for amplification in bacteria)

43
Q

What is the general concept of gene therapy?

A

Use patients as the host for protein production

44
Q

How could gene therapy be used to permanently treat type 1 diabetes?

A

Turn liver cells into insulin making machines

Biochemistry (3 decks)

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