Gene Tech_Set 1

A2 Biology
By Akram Sir
3 During an immune response, only B-lymphocytes with receptors that are specific to the antigens present are activated.
Activation occurs when an antigen binds to a receptor of a B-lymphocyte.Activated B-lymphocytes grow in size and then divide
by mitosis. Many further mitotic cell divisions occur, increasing the number of B-lymphocytes with receptors specific to the antigen
. Eventually, cells produced in this process will develop into either plasma cells that secrete antibodies or memory B-cells.
Fig. 3.1 is a summary of B-lymphocyte activation and the events that follow.
 The development of plasma cells and memory B-cells in this process
depends on transcription factors.
 
 
(a) Explain the role of transcription factors in gene expression in eukaryotic
cells.

1 binds to DNA ;
2 at, promoter / enhancer ;
3 allows, RNA polymerase / other transcription factors, to bind (to, DNA /
gene / promoter) ;
4 regulates / initiates / inhibits, gene expression / transcription ;
A switches genes, on / off
5 (so that genes are expressed) at the correct time / in the correct context / in
the correct cell type / in the correct order ;

Transcription factors are protein molecules which can bind to DNA at

promoter region which is upstream to the specific gene and allows binding
of RNA polymerase for the formation of mRNA which switch on the gene
expression.
 
(a) Transcription factors are proteins. Genes that code for proteins can
become mutated.
Describe how different types of gene mutation can cause changes in the
protein that is synthesized.
1 BLIMP-1, is not synthesised / is non-functional / has changed function ;
2 expression of the c-Myc gene, is not reduced / continues ;
3 synthesis / concentration, of c-Myc (protein), is maintained / increases ;
4 (B-lymphocytes continue to) divide by mitosis / proliferate ;
A clonal expansion
5 differentiation / specialisation, prevented ;
c) BLIMP-1 is a transcription factor that is essential for the development of
plasma cells and memory B-cells in the process shown in Fig. 3.1.
BLIMP-1 reduces the synthesis of c-Myc in B-lymphocytes. c-Myc is a
protein that is required for the mitotic cell cycle to continue.
 
Suggest and explain how a mutation in the gene coding for BLIMP-1 can
prevent the development of plasma cells and memory B-cells.

1 base substitution / mis-sense mutation ;

2 changes, triplet / codon ;
3 base, deletion / insertion ;
4 (results in) frame shift / description ;
5 change in, protein primary structure / amino acid sequence ;
6 protein folds incorrectly / changes tertiary or 3-D structure ;
7 changes protein function / prevents protein function / makes protein unstable;
8 idea of new STOP codon ;
9 only, short / first part of / no, protein is produced
(d) Microarrays can be used to analyse the effect of transcription factors,
such as BLIMP-1, on gene expression.
 
(i) Describe how a microarray is used in the study of gene expression.

1 mRNA extracted from cells (of interest) ;

2 (mRNA) used (as template) to synthesise cDNA ;
3 cDNA is tagged with a fluorescent dye ;
4 (fluorescent) cDNA, binds to / hybridises with, probe ;
5 each probe is unique to a different gene / AW ;
6 fluorescence indicates gene is expressed ;
7 AVP ; e.g. detail of probe / use of UV light
(ii) BCL6 is another transcription factor found in B-lymphocytes. The effect of
BCL6 on gene expression was compared in two samples of B-lymphocytes.
 
 Sample 1 consisted of B-lymphocytes that were producing BCL6.
 Sample 2 consisted of B-lymphocytes that were not producing BCL6.
 
Suggest why a microarray is suitable for identifying the function of the
transcription factor BCL6 in these two samples.

1 BCL6 / transcription factors, regulate expression of genes ;

2 (microarray) can detect expression of genes ;
3 idea of difference in, fluorescence / gene expression, between
two samples ;
4 difference in gene expression indicates regulation by BCL6 ;
(b) Transcription factors are proteins. Genes that code for proteins can
become mutated. Describe how different types of gene mutation can cause
changes in the protein that is synthesized.

1 BCL6 / transcription factors, regulate expression of genes ;

2 (microarray) can detect expression of genes ;
3 idea of difference in, fluorescence / gene expression, between two samples ;
4 difference in gene expression indicates regulation by BCL6 ;
1 repeat treatment causes, bigger / more rapid, immune response ;
2 (so) target cells (expressing new protein) short-lived / AW ;
3 decrease in (trans)gene expression on re-administration ;
4 AVP ; e.g. ref. to antibody binding / antibodies attract phagocytes
any four from:
naked DNA
1 has to be injected into target cell / lack of organ-specific delivery ;
2 low efficiency of cellular uptake ;
3 rapidly broken down ;

viruses
4 small packaging capacity / only small amount of DNA can be carried ;
5 low probability of integration (into host genome) ;
6 cause mutations in host DNA / (gene) insertion disrupts gene function / insertional
mutagenesis ;

liposomes
7 low ability to, add DNA / genes, into target cells (genome) / low transduction
efficiency ;
1 red blood cells had ‘self’ antigens ;
2 not recognised as foreign
(red blood cell) has no nucleus so no, DNA / transcription / translation, of
targeting proteins ; ora