SECTION B: STRUCTURED QUESTIONS

Please write down your responses on writing papers.

N09/P2/2
Fig. 1.1 shows the arrangement of the bacterial chromosome and F plasmid in a bacterial cell.

(a)

State two ways in which a plasmid such as the F plasmid differs from the bacterial
chromosome.
1. The plasmid is smaller than bacterial chromosome;
containing fewer genes than bacterial chromosome;
2. Genes found on plasmid may code for antibiotic resistance or pilus formation;
while genes on bacterial chromosome code for enzymes needed for cell
metabolism;

(b)
(i)

[2]

Outline the processes of

bacterial conjugation and
- A F+ donor cell has sex pili that bind to specific receptors on a F- recipient cell;
- Upon contact, the sex pilus retracts to bring the F- cell closer to F+ cell and a mating
bridge forms between the 2 cells;
- One strand of ds plasmid is cleaved by enz at origin of transfer while other strand
moves into recipient cell;

11

- As transfer continues, donor F plasmid replicates using ss F plasmid as

template for synthesis of new strand in both donor and recipient cells;
(ii)

[2]

bacterial transformation.
- free ds exogenous DNA binds to DNA binding complexes on cell wall of competent
bacterium;
- one strand degraded, other strand taken up into cell;
- homologous recombination occurs which results in a heteroduplex;
- where one strand has original allele while the other has allele from exogenous
DNA;

(c)

[2]

Suggest two potential benefits of conjugation for the recipient bacteria.

- Development of antibiotic resistance;;
- Gain ability to make use of a new metabolite;;

(d)

[2]

Outline the main stages in bacterial transduction.

- lytic bacteriophate infects a bacteria and upon penetration, bacteriophage enz lyse
bacterial genome into fragments;
- during assembly of progeny phages, some fragments of lysed bacterial genome
mistakenly get incorporated into capsid of progeny bacteriophages;
- progeny phage carrying fragment of bacterial genome infects another bacterial cell;
- during penetration, fragment of bacterial genome injected into new bacterial host;
- transduced bacterial genome incorporated into genome of new host;
- via homologous recombination;

[3]

[Total:11]
SECTION C: FREE RESPONSE QUESTIONS
Please write down your responses on writing papers.
Question 1
N07/P2/Q9
Describe how the prokaryote, E.coli, is able to respond to varying concentrations of
lactose.
[6]
(optional)
- (Lactose metabolism is controlled by enzymes coded for by the lac operon)
- The structural genes involved are lacZ, lacY and lacA which code for beta-galactosidase,
permease and transacetylase respectively;;

12

- (These structural genes are under the control of a single promoter and operator);
- (Therefore, their expression is coordinately regulated depending on the level of lactose in
the cell);
- When lactose levels are low, the lac repressor protein is active;
- and bound to the operator of the operon;
- This prevents binding of RNA polymerase to the promoter;
- and transcription of structural genes is switched off;
- When lactose levels are high, lactose enters the cell (via permease);
- and is converted to allolactose;
- Allolactose acts as an inducer that binds to the lac repressor protein;
- inactivating the protein;
- RNA polymerase can now bind/ access the promoter;
- and transcription of enzymes involved in lactose metabolism is activated;
Question 2
N07/P2/Q9
(a)

Describe the main structural features of the lac operon.

1.
2.
3.
4.

Diagram of lac operon

Lac operon consist of 3 structural genes involved in lactose metabolism;
lac Z coding for -galactosidase;
which breaks down lactose into galactose and glucose + converts some
lactose into allolactose;
5. lac Y coding for -galactoside permease;
6. that transports lactose into bacteria cytosol from surroundings;
7. lac A coding for -galactoside transacetylase;
8. which may be involved in the removal of metabolic toxins from lactose
breakdown;
9. All 3 structural genes under control of same promoter and operator;
10. Promoter contains 2 binding sites;
11. one for RNA pol;
12. and other for cAMP-CAP complex;
13. Operator contains binding site for lac I repressor prot;
14. which regulates transcription of the 3 structural genes;
15. by controlling the access of RNA pol to promoter;
16. lac I gene and own promoter is located upstream from promoter of lac
operon;
17. lac I codes for repressor protein;

13

[8]

(b)

Explain how the presence of lactose in the growth medium switches on the lac
operon.

[8]

1. in absence of lactose, lac I repressor prot is active;

2. binds to operator site of lac operon;
3. preventing access of RNA pol to promoter;
4. hence no transcription of structural genes occurs and lac operon is OFF;
5. in presence of lactose, lactose enters bacteria via transmemb prot permease;
6. basal levels of enz -galactosidase in cell;
7. will break down lactose into glucose and galactose;
8. and converts lactose into its isomer, allolactose;
9. allolactose is the inducer of the lac operon;
10. binds to free and bound lac I repressor prot in cytoplasm and operator
respectively;
11. causes conformational change in repressor prot;
12. such that it cannot bind to operator;
13. RNA pol able to access promoter;
14. proceeds to transcribe the structural genes into a polycistronic mRNA and lac
operon is ON;
15. mRNA will then be translated by free ribosomes to produce the enzymes for
lactose metabolism;
16. lac operon is thus a negatively controlled inducible operon;

(c)

Suggest why operons are necessary in bacteria.

1.
2.
3.
4.
5.

[4]

Operons are a linear arrangement of genes and its regulatory sites;

These genes which are clustered together;
are under the control of a common promoter;
and encode proteins which execute similar functions;
hence allowing a common mode of regulation/ coordinated control of gene
expression;
6. in which proteins performing similar functions are turned on together;
7. Operons help bacteria maximize resource utilization/ conserve energy;
8. by not transcribing genes whose products are not needed;

14

EXTRA/SUPPLEMENTARY QUESTIONS
Question 1

Fig. 1.1 shows a prokaryotic cell.

D

C
B

A
E

A
F

Fig. 1.1
(a)

(bi)

Name structures A H.
A flagella;

B plasma membrane;

C ribosomes;

D pili;

E plasmid;

F dsDNA/ bacterial chromosome;

G cytoplasm/ cytosol;

H capsule/ cell wall;

[4]

Name the region in the cell where bacterial chromosome can be found.
nucleoid;;
[1]

(bii) Explain how the chromosome is packed into the region stated in (b)(i).
- bacterial chrom is circular;

- negative supercoiling;
- highly compacted;
- due to formation of loop domains;
- association with histone-like proteins;
- for DNA to bend for compaction;
(c)

[3]

State why plasmid replication is independent of the chromosomal DNA.

- plasmid is circular extrachromosomal dsDNA;
- has own origin of replication;

[1]

15

(d)

Explain the process employed by bacteria during colonization.

- binary fission;
- division of one bacteria cell to produce 2 daughter cells;
- parental chromosome acts as template for replication of daughter DNA;
- daughter DNA attached to memb at site adj to parental attachment site;
- bacterial cell elongates during DNA replication;
- after DNA fully replicated and bacteria has doubled in size;
- plasma membrane invaginates, dividing parent cell into 2 daughter cells;
- cell wall material deposited above membrane to form new cell wall;

(e)

[4]

Name an organelle in eukaryotic cells that has similar properties as a bacterial cell.
Mitochondrion / chloroplast;;
(because it contains own DNA)

[1]

[Total:14]

Question 2
(a)

Distinguish between structural and regulatory genes in a prokaryotic genome, and give an
example for each.
- structural genes a region of DNA that codes for a protein or RNA molecule that
forms part of a structure or has an enzymatic function;;
- e.g. lac Z gene;
- regulatory genes a region of DNA that codes for specific prot that regulates
expression of structural genes;;
- e.g. lac I gene;

[3]

16

(b)

Fig. 1 shows a simple lac operon when there is no lactose.

Q
R

Fig. 2

Label the parts P, Q, R, S.

(c)

active lac repressor;

promoter;

operator;

mRNA;

[2]

Describe the negative control mechanism of lac operon in the presence of lactose and
absence of glucose.
- in presence of lactose and absence of glucose, basal levels of -galactosidase
within bacteria converts lactose to allolactose;
- allolactose binds to active repressor causing it to change conformation;
- repressor can no longer bind to operator;
- RNA pol can access promoter and proceeds to structural genes lac Z, Y, A;
- since repressor is inactive in presence of inducer allolactose resulting in activation
of transcription;
- and repressor is active in absence of inducer allolactose resulting in inhibition
of transcription, lac operon is said to be under NEGAIVE control;

(d)

State and describe one process through which genetic diversity can be achieved in bacteria.
Any 1 of the following with correct descriptions:

(1)

[3]

- conjugation;

17

- F + donor has sex pili that binds to specific receptors on F - cell;

- upon contact, sex pilus retracts to bring F- cells closer to F+ cell;
- forming mating bridge which provides physical contact b/w cells;
- one strand of ds plasmid cleaved by enz at oriT and ss F plasmid moves into
recipient cell;
- donor F plasmid replicates using ss F plasmid as template for synthesis of new
strand in both donor and recipient cells;
(2)

transduction;
- a bacteriophage transfers bacterial genes from one host cell to another;
- generalized erroneous packaging of lysed host genome into capsid during
assembly of viral progeny;
- specialized erroneous excision of specific part of bacterial genome near to
prophage during lytic induction from lysogenic cycle;
- viral progeny containing bacterial genome re-infect other host cells;
- recombination b/w homologous regions of host genome gives rise to recombinant
bacteria;

(3)

transformation;
- uptake of exogenous/ foreign DNA into cell;
- dsDNA binds to specific cell surface receptors of competent bacterium;
- 1 DNA strand degraded, other strand enters bacteria;
- recombination b/w homologous regions of bacterial chrom and exogenous DNA;
- heteroduplex forms, where 1 strand has original allele, other strand has alleles from
exogenous DNA;
[3]
[Total:11]

Question 3

Fig. 3.1 shows two bacteria in the process of transferring genes.

A

Fig. 3.1
18

(a)

Name and briefly describe the role of A

- A is a conjugation bridge( sex pilus);
- allows physical contact between 2 bacterial cells;
- enables transfer of F plasmid from F+ cell to F- cell;
- conversion of bacteria from sterile to fertile;

(bi)

[2]

Identify and describe the process shown above.

- process conjugation;;
- F+ donor cell has sex pili;
- which bind to specific receptors on F- cells;
- upon contact, sex pilus retracts to bring F- cell closer to F+ cell;
- a mating bridge is formed between the 2 cells to allow DNA transfer;
- 1 strand of ds plasmid cleaved by enz at origin of transfer;
- ss F plasmid moves into recipient cell;
- as transfer continues, donor F plasmid replicates using ss F plasmid as
template;
- synthesizing new strand in both donor and recipient cells;
- transferred plasmid circularizes in recipient cell;
- recipient cell now becomes a F+ donor cell and has the ability to form sex
pili;

[6]

(bii) Suggest the importance of this process.

- allows transfer of resistance factors among bacteria;

- increase genetic variation among bacteria;

(ci)

[1]

High frequency of recombinant (Hfr) cells are capable of transferring chromosomal DNA
to a F- cell. Describe how this may occur.
- F plasmid integrated into chromosomal DNA;
- F plasmid contains origin of transfer;
- during conjugation, ssDNA thats being transferred includes part of bacterial
genome DNA;
- chromosomal DNA of Hfr bacteria gets transferred to F- strain;

19

- transfer usually incomplete, not entire F plasmid transferred;

- F- strain remains sterile;
- transferred portion may undergo recombination with F- genome;
- possess phenotypic traits of the F+ strain;

(cii)

[4]

Compare and contrast between a Hfr and a F+ bacteria cell.

Similarities

Both produces sex pili for transfer of DNA;

Attach to F- cell;
Differences

Feature

F+

Hfr

Fertility (F) factor

Integrated into
chromosomal DNA

Extrachromosomal
DNA

Transfer of DNA

Transferred part of
chromosomal DNA
with genes from
bacteria to F- cell

Only F plasmid
transferred to F- cell

Transfer of F factor

Part of F factor
transferred

Whole factor
transferred

State of F- cell after

transfer

sterile

fertile

[5]

(d)

Compare and contrast the process in (bi) with transduction and transformation.
Similarities

Allows gene transfer between bacteria;

Might result in recombinant genome/ increase genetic variation among bacteria;

Differences

Feature

Conjugation

Medium for gene Bacteria. Both

transfer
needs to have
physical contact

20

Transduction
Bacteriophages
during
erroneous
excision of

Transformation
From
surrounding
medium where
lysed bacteria

via sex pili

genome/
packaging of
genome into
capsid

genome is free

Fragment
transferred

F+ transfer
whole F plasmid
to F- cell

Part of the
bacteriophageinfected genome
is transferred to
another bacteria

Usually small
fragment since
floating
fragments would
have been
degraded prior
entry into cell

Process

Active via
formation of sex
pili from F+/ Hfr
cell

Active via
transfer by
bacteriophages

Passive, taken
up from
surrounding
medium
[7]

[Total:25]

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