Question
There are many different forms of Vibrio cholerae, a bacterium that is found naturally in aquatic environments. The bacterium is motile (can move) and uses a cell structure known as a flagellum to allow it to move through water.
Fig. 4.1 is a drawing of four cells of one form of V. cholerae.
flagellum
X Y
cell wall
Fig. 4.1
Two main forms of V. cholerae, O1 and O139, are able to colonise the small intestine and cause cholera. These two forms are able to produce a toxin, choleragen, which causes the symptoms of diarrhoeal disease. Mutant V. cholerae that lack flagella are less able to cause disease.
(a) The magnification of the diagram shown in Fig. 4.1 is ×32 000.
Calculate the actual width X–Y in Fig. 4.1 in nanometres (nm) and give your answer to the nearest 10 nm.
Complete Fig. 4.2 to show the formula you will use to make your calculation.
actual
= width
Fig. 4.2
answer = …… nm [2]
(b) State the term used to describe disease-causing organisms, such as the bacterium
V. cholerae.
…… [1]
(c) Outline one way in which an uninfected person may become infected by V. cholerae.
…… [1]
(d) Choleragen is produced after V. cholerae has penetrated (passed through) the mucus lining and attached to intestinal epithelial cells.
Choleragen is composed of two subunits:
• subunit A consists of one polypeptide
• subunit B consists of five identical polypeptides
• the polypeptide in subunit A is different from the polypeptides in subunit B.
Two genes, ctxA and ctxB, are needed to produce choleragen. Only one strand of the DNA forming gene ctxA is involved in the production of subunit A. Only one strand of the DNA forming gene ctxB is involved in the production of subunit B.
Explain why only one strand of the DNA of each gene is involved in the production of the subunits.
…… [3]
Monoclonal antibodies (mAbs) can be designed to act against components of the cell wall of V. cholerae. The cell wall has an outer membrane with lipopolysaccharide (LPS) molecules, shown in Fig. 4.3.
O-polysaccharide
outer lipopolysaccharide membrane (LPS) core polysaccharide cell wall
lipid A
fluid space
peptidoglycan layer
Fig. 4.3
The core polysaccharide and the lipid A components of the LPS molecules are the same in
V. cholerae O1 and V. cholerae O139. However they have different O-polysaccharides.
There are also different types of V. cholerae O1 and these have different O-polysaccharides.
(e) Laboratory tests were carried out using two different monoclonal antibodies that had been designed and produced to act against the LPS of bacterial cultures of V. cholerae O1:
• mAb 2D6 acts against the O-polysaccharide
• mAb ZAC-3 acts against the core polysaccharide and lipid A components.
(i) Explain why the mAb ZAC-3 produced against the core polysaccharide and lipid A components will not act against the O-polysaccharide of the LPS molecules.
…… [2]
(ii) The results of the tests showed that both mAbs were effective in causing agglutination
(clumping) of bacteria and in preventing their motility. This suggests they may be useful for preventing cholera and for treating the disease.
Discuss whether mAb 2D6 and mAb ZAC-3 may be useful for preventing cholera and for treating the disease.
…… [4]
[Total: 13]
Mark scheme
4(a) image length ; A magnification triangle 2
magnification
250 (nm) ; (8 mm)
A 220 (for 7 mm)
A 230 (for 7.5 mm)
A 270 (for 8.5 mm)
A 280 (for 9 mm)
4(b) pathogen ; 1
4(c) ingests / drinks / AW, contaminated water / water containing the pathogen / food washed in contaminated 1
water / contaminated food ;
A examples of accidental ingestion e.g. swimming in contaminated rivers
A faecal / oral, transmission
A bacteria / Vibrio
R if described as virus
4(d) A mRNA or messenger RNA 3
any three from:
(one strand only needed) to form mRNA / mRNA is single-stranded ;
(m)RNA is, used / needed, to produce, subunit (s) / polypeptide(s) ;
(only) one strand (of DNA) is the, template / transcribed, strand ;
idea that (complementary) copying / transcribing, other DNA strand would not result in, desired / AW, mRNA / polypeptide ;
4(e)(i) any two from: 2
I receptor I active site
(mAbs) specific / different, antigen binding sites / binding sites for antigen ;
A (each) mAb binds to a, specific / particular, antigen
R if antibody described as an enzyme
(each mAb has) specific / different, tertiary structure / variable region(s) / primary structure / sequence of amino acids ;
binding site and antigen have complementary shapes
or
ZAC-3 complementary shape to core polysaccharide and lipid A
or
2D6 complementary shape to O-polysaccharide ;
4(e)(ii) any four from: 4
yes
general points
(agglutination / motility prevented, so)
bacteria less able to, penetrate mucus / attach to intestinal epithelial cells / colonise intestine / AW ;
A idea that fewer bacteria able to, attach / colonise intestine
less / no, choleragen / toxin, released ;
bacteria passed out in faeces not able to cause disease in others / AW ;
ref. to phagocytosis more effective ;
e.g. macrophages stimulated to carry out phagocytosis
prevention / treatment
(to prevent disease) needs to be given, at early stages / before colonising (intestine) ;
passive immunity / passive vaccine ;
must be in context of treatment or immediate protection
idea that in addition to immune response (so increased effect) ;
e.g. acts beside, immune system / immune system cells
acts before the immune response can become effective
ref. to quicker recovery (if a person has cholera) ;
useful when, there is antibiotic resistance / antibiotics cannot be given ;
specific mAb
mAb ZAC-3 may be more effective for cholera caused by, wider range of V. cholerae / AW;
mAb ZAC-3 may be useful if exact form of V. cholerae not known ;
mAb 2D6, needs to be targeted against specific V. cholerae forms / may not be effective against other V. cholerae forms ;
A not all forms of V. cholerae were tested in lab
4(e)(ii) AVP ; e.g.
(only) short-term protection / not active immunity / not long term
ref. difficulty in delivering mAbs to intestine
e.g. may be, digested / destroyed, as pass through gut
need to find a way to get mAbs to the intestine
if given intravenously need to pass through to gut lumen
may be able to prevent multiplication of bacteria
mAbs have been used successfully for other diseases