MCB2011 - Molecular biology and the cell - S1 2025
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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.
Identify the donor and target membranes for the secretory vesicle. (2
marks)
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.
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
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.
Examine the western blotting results shown in the image above (A, B, C). Which result would you expect to obtain for this experiment?
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)
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.
Identify the donor and target membranes for the secretory vesicle. (2
marks)
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:
The process depicted in the electron micrograph shown above is classified as:
A researcher decides to compare the promoter regions of . 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).
Use the drop-down boxes to indicate which amplified promoter region ( or
Lane 1:
Lane 2:
Lane 3:
CONTINUED...
The ecologists identify a putative protein-encoding gene ( ) 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.
What was likely omitted from the sequencing reaction? Be specific in your response and explain your reasoning. (2-3 sentences) (2 marks)
The researcher decides to design a polymerase chain reaction (PCR) assay to screen HCM patients for the risk allele. To determine if a patient is homozygous for the risk allele, homozygous for the reference allele, or heterozygous (i.e. carries one copy of each allele), the assay must simultaneously amplify and measure the presence of both alleles (risk and reference).
Which approach would you recommend for the assay? (1 mark)
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