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MCB2011 - Molecular biology and the cell - S1 2025

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During the phylotypic stage, vertebrate embryos from different species ______________________________.

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The researcher

captured the electron micrograph image below which shows the release of albumin

from a secretory vesicle within a hepatocyte into the extracellular

environment.

Image failed to load

Identify the donor and target membranes for the secretory vesicle. (2

marks)

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The import of acyl-CoA dehydrogenase into the mitochondrion is defective

in cell line HEP-M1. The researcher used subcellular fractionation and western

blotting to detect acyl-CoA dehydrogenase in different subcellular

compartments. The western blotting results for normal healthy hepatocytes and the

HEP-M1 cell line are summarised in the figure below.

Image failed to load

Which of the

following mutations (A, B, C) is most likely present in the HEP-M1 cell line?

Use the western blotting results and your knowledge of the mitochondrial

protein import process to justify your response. (2-3 sentences) (2 marks)

A. The acyl-CoA

dehydrogenase mitochondrial signal sequence is mutated

B. The

receptor protein in the TOM complex is mutated

C. The

translocation channel in the TIM23 complex is mutated

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A researcher extracts protein from cultured hepatocytes (i.e. liver

cells) and treats the protein sample with sodium dodecyl sulphate (SDS) and

β-mercaptoethanol (BME). The protein sample is loaded onto a polyacrylamide gel

and a western blot is performed as per the steps described in the relevant online

learning module (OLM 6.2). The researcher uses a primary antibody that specifically

binds to the N-terminal region of acyl-CoA dehydrogenase.

Image failed to load

Examine the

western blotting results shown in the image above (A, B, C). Which result would

you expect to obtain for this experiment?

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The researcher

suspects that low serum albumin levels in affected patients are not directly

caused by acyl-CoA dehydrogenase defects, but are a secondary consequence of

broader metabolic stress. To explore how acyl-CoA dehydrogenase defects alter

cellular protein expression, the researcher extracts total protein from normal healthy

hepatocytes and a mutant hepatocyte line (HEP-M2).

Which proteomic

technique would you use to compare the protein profiles of the two samples? Explain

why your chosen technique is suitable for this specific application. (2-3

sentences) (2 marks)

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The researcher

captured the electron micrograph image below which shows the release of albumin

from a secretory vesicle within a hepatocyte into the extracellular

environment.

Image failed to load

Identify the donor and target membranes for the secretory vesicle. (2

marks)

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A signal sequence at the N-terminus of the albumin protein directs it to

the correct subcellular compartment following translation. The albumin signal

sequence likely contains:

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The process depicted in the electron micrograph shown above is classified as:

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A researcher decides

to compare the promoter regions of

AlpA, ACTB and ACTC1

.

The researcher uses traditional PCR to amplify the promoter regions and

visualises the PCR products using agarose gel electrophoresis. The length of

the promoter region of each gene is summarised in the table below (A). The agarose

gel electrophoresis results are also shown below (B).

Image failed to load

Use the

drop-down boxes to indicate which amplified promoter region (

AlpA, ACTB

or

ACTC1) is present in each lane of the agarose gel. (0.33 marks each, 1 mark total)

Lane 1:

Lane 2:

Lane 3:

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CONTINUED...

The ecologists

identify a putative protein-encoding gene (

AlpA

) in the genome of the

novel organism and use Sanger sequencing to determine the gene sequence. The sequencing

results are shown in the electropherogram below. Unfortunately, an error was

made setting up the sequencing reaction. As a result, the reaction terminated prematurely

at the ninth nucleotide.

Image failed to load

What was likely

omitted from the sequencing reaction? Be specific in your response and explain

your reasoning. (2-3 sentences) (2 marks)

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