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Vadim26 [7]
3 years ago
14

You are performing a biochemical purification of enzymes involved in DNA replication. You have purified the replisome. You wish

to purify the primosome. You perform further separation techniques on your purified replisome. How will you test to determine that you have purified the primosome
Biology
1 answer:
algol [13]3 years ago
6 0

Confirm that the sample has both helicase and primase activities, but not the ability to synthesize DNA

  • The replisome is a multiprotein-RNA complex that is required for the replication of DNA. The replisome consists of different proteins which include:

            1. Helicase enzymes that unwind and separate the DNA strand

            2. Replication factor C (RFC), a DNA-dependent ATP that acts as an activator of DNA polymerase.

           3. Proliferating cell nuclear antigen (PCNA)

           4. DNA gyrase/topoisomerase that cuts and reseals DNA strands, which is essential for DNA synthesis.

           5. Primase enzyme that synthesizes short RNA fragments called primers.

          6. DNA polymerase III, an enzyme that catalyzes the polymerization of deoxyribonucleotides into a DNA strand.

           7. DNA ligases enzymes that form phosphodiester bonds at a single-strand break in DNA

  • A primosome is a multiprotein complex responsible for creating RNA primers on single-stranded DNA (ssDNA) during the replication process.
  • The primosome is composed of different primase and helicase enzymes. In<em> Escherichia coli</em>, it consists of PriA helicase, PriB, PriC, DnaB helicase, DnaG (encoded by the dnaG gene), DnaC and DnaT primase.

Learn more in:

brainly.com/question/14674568?referrer=searchResults

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In Drosophila melanogaster, vestigial wings (vg) is recessive to normal wings (vg+), black body (b) is recessive to gray body (b
yan [13]

Correct progeny phenotype:

  • 1779 vestigial wings, black body and purple eyes, vg b pr  
  • 1665 normal wings, a grey body and red eyes, vg+ b+ pr+
  • 252 normal wings, an black body and purple eyes, vg+ b pr
  • 241 vestigial wings, a gray body and red eyes, vg b+ pr+
  • 131 normal wings, an black body and red eyes, vg +b pr+
  • 118 vestigial wings, a gray body and purple eyes, vg b+ pr
  • 13 vestigial wings, an black body and red eyes, vg b pr+
  • 9 normal wings, a gray body and purple eyes, vg+ b+ pr

Answer:

  • The order of these genes is vg --- pr --- b
  • Map distances between the genes vg/pr = 12.2 MU
  • Map distance between the genes pr/b = 6.4 MU
  • Map distances between the genes vg/b = 18.6 MU

Explanation:

We know that

•Normal wings expressed by vg+ is dominant over vestigial wings, vg

•Gray body b+ is dominant over black body

•Red eyes, pr+, is dominant over purple ayes, pr

We have the number of descendants of each phenotype product of the tri-hybrid cross.

•1779 vestigial wings, black body and purple eyes vg b pr  

•1665 normal wings, a grey body and red eyes vg+ b+ pr+

•252 normal wings, an black body and purple eyes vg+ b pr

•241 vestigial wings, a gray body and red eyes vg b+ pr+

•131 normal wings, an black body and red eyes vg +b pr+

• 118 vestigial wings, a gray body and purple eyes vg b+ pr

•13 vestigial wings, an black body and red eyes vg b pr+

• 9 normal wings, a gray body and purple eyes vg+ b+ pr

The total number of individuals is 4208.

In a tri-hybrid cross, it can occur that the three genes assort independently or that two of them are linked and the third not, or that the three genes are linked. In this example, in particular, the three genes are linked on the same chromosome.

Knowing that the genes are linked, we can calculate genetic distances between them. First, we need to know their order in the chromosome, and to do so, we need to compare the genotypes of the parental gametes with the ones of the double recombinants. We can recognize the parental gametes in the descendants because their phenotypes are the most frequent, while the double recombinants are the less frequent. So:

<u>Parental)</u>

  • 1779 vestigial wings, black body and purple eyes vg b pr  
  • 1665 normal wings, a grey body and red eyes vg+ b+ pr+

<u>Double recombinant)</u>

  • 13 vestigial wings, an black body and red eyes vg b pr+
  • 9 normal wings, a gray body and purple eyes vg+ b+ pr

<u>Simple recombinant)</u>

  • 252 normal wings, an black body and purple eyes vg+ b pr
  • 241 vestigial wings, a gray body and red eyes vg b+ pr+
  • 131 normal wings, an black body and red eyes vg +b pr+
  • 118 vestigial wings, a gray body and purple eyes vg b+ pr

Comparing parental with the double recombinants we will realize that between  

  • vg b pr (parental)
  • vg b pr+ (double recombinant)

and  

  • vg+ b+ pr+ (Parental)
  • vg+ b+ pr (double recombinant)

They only change in the position of the alleles pr/pr+. This suggests that the position of the gene pr is in the middle of the other two genes, vg and b, because in a double recombinant only the central gene changes position in the chromatid.  

So, the order of the genes is:

---- vg ---- pr -----b ----

Now we will call Region I to the area between vg and pr and Region II to the area between pr and b.

Once established the order of the genes we can calculate distances between them, and we will do it from the central gene to the genes on each side. First We will calculate the recombination frequencies, and we will do it by region. We will call P1 to the recombination frequency between vg and pr genes, and P2 to the recombination frequency between pr and b.

P1 = (R + DR) / N

P2 = (R + DR)/ N

Where: R is the number of simple recombinants in each region, DR is the number of double recombinants in each region, and N is the total number of individuals.  

So:

Parental)

• 1779 vestigial wings, black body and purple eyes vg pr b  

• 1665 normal wings, a grey body and red eyes vg+ pr+ b+  

Double recombinant)

• 13  vg pr+ b  

• 9  vg+ pr b+  

Simple recombinant)

• 252  vg+ pr b  

• 241 vg pr+ b+  

• 131  vg+ pr+ b  

• 118  vg pr b+  

P1 = (R + DR) / N

P1 = (252+241+13+9)/4208

P1 = 515/4208

P1 = 0.122

P2= (R + DR) / N

P2 = (131+118+13+9)/4208

P2 = 271/4208

P2 = 0.064

Now, to calculate the recombination frequency between the two extreme genes, vg and b, we can just perform addition or a sum:

P1 + P2= Pt

0.122 + 0.064 = Pt

0.186=Pt

The genetic distance will result from multiplying that frequency by 100 and expressing it in map units (MU). One centiMorgan (cM) equals one map unit (MU).  

The map unit is the distance between the pair of genes. Every 100 meiotic products, one of them results in a recombinant product. Now we must multiply each recombination frequency by 100 to get the genetic distance in map units:

GD1= P1 x 100 = 0.122 x 100 = 12.2 MU

GD2= P2 x 100 = 0.064 x 100 = 6.4 MU

GD3=Pt x 100 = 0.186 x 100 = 18.6 MU

---- vg ---------------------- pr ---------------------b ----

                    R1                                 R2

-----vg----12.2MU---------pr—

                                   ----pr--------6.4 MU----b—

-----vg ----------------18.6 MU--------------------b----

                                   

3 0
3 years ago
You have a large bottle of 2% saline solution in the lab (2% NaCl, which is sodium chloride). You need 5 ml of 0.1% saline for a
zvonat [6]

Answer:

volume of 2% saline (V₁) = 0.25ml

volume of water = 4.75ml

Explanation:

The requirement in the question is to obtain 0.1% of NaCl from 2% NaCl, and the process is dilution. The general dilution equation is guven by:

C₁V₁ = C₂V₂

where:

C₁ = initial concentration = 2%

V₁ = initial volume = ?

C₂ = final concentration = 0.1%

V₂ = final volume = 5ml

C₁V₁ = C₂V₂

calculating for V₁

∴ V₁ = (C₂V₂) ÷ C₁

= (0.1% × 5) ÷ 2%

V₁ = 0.25ml

Now, let us calculate the volume of water needed:

V₁ + Volume of water = V₂ (volume of 0.1% NaCl)

0.25 + volume of water = 5

volume of water = 5 - 0.25

volume of water = 4.75ml

8 0
3 years ago
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