GENOME PROJECTS AND MAKING DNA FRAGMENTS Flashcards

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1
Q

Genome

A

a genome is the entire set of dna including all genes in an organism

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2
Q

how does gene sequencing occurs

A

it can only be done in fragments of dna.

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3
Q

proteome

A

the proteome of an organism is all the protein made by it.
- proteome can be determined by dna sequencing of their genome

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4
Q

translating genome in complex organism

A

-complex organism contains a large section of non coding dna - in addition to that they also contain regulatory genes
- difficult to translate the genome into the proteome as hard to find the bits that codes fro proteins.

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5
Q

recombinant dna technology

A

recombinant dna technology involves transferring a fragment of dna from one organism to another.

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6
Q

dna fragments

A

inorder to transfer gene, we first need to get a dna fragment which consists of the target gene.

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7
Q

how can you make dna fragments

A
  1. using reverse transcriptase
  2. using restriction endonuclease enzymes
  3. using a gene machine
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8
Q

sequencing a genome

A

working out dna base sequence for all dna in a cell

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9
Q

reverse transcriptase

A
  • reversing the transcription process ( mrna converted into dna)
  • reverse transcriptase makes dna copies from mrna
  • first mrna is isolated from the cells that contains a large amount of mrna for protein
  • mrna is then mixed with dna nucleotides and reverse transcriptase - in a way that mrna acts as a template to synthesize a new dna strand.
  • reverse transcriptase joins dna nucleotides with complentary mrna bases.
  • a single stranded dna called cdna is produced.
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10
Q

advantage of cdna

A
  • cdna is intron free as it is based on mrna template.
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11
Q

restriction endonuclease enzyme

A
  • some sections of dna have palindromic sequences of nucleotides ( anti parallel base pairs that read the same in opposite directions)
  • restriction endonuclease are the enzymes that recognise specific palindromic sequence also called recognition sequence and cut dna at that point.
  • different restriction endonuclease cut at different recognition sequence because the shape of the recognition sequence is complementary to active site of the enzyme.
  • the dna sample is incubated with specific restriction endonuclease which cuts the dna fragment via a hydrolysis reaction.
  • sometimes the cut leaves sticky ends
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12
Q

sticky ends

A

sticky ends are the small tails of unpaired dna bases at the end of the fragment.
- sticky ends can be used to bind dna fragment with another piece of dna that has a sticky end with complementary sequences.

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13
Q

gene machine

A
  • fragments of dna can be synthesized from scratch
  • dna fragments are made in a lab
  • first amino acid sequence is identifed for mrna and dna sequence .
  • dna sequence is entered into computer for bio safety
  • first nucleotide in the sequence is attatched to some sort of support eg beads
  • nucleotides are then added step by step in correct prder which also includes adding protecting groups
  • short sections of dna arecreated called oligo nucleotides , roughly 20 nucleotides long.
  • once complete they are broken off from the support and all protecting groups are removed. oligonucleotides are joined together to create longer dna fragments.
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14
Q

why do we add protecting groups while making dna in a gene machine

A
  • to ensure nucleotides are joined at the right place, and to prevent unwanted branching.
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