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School of Biological Sciences Year 1 Semester 2

2015/2016
BOI 116/4- GENETICS
Report of Mini project 2: Drosophila
Melanogaster Genetics
Name: Tham Khai Xin
Matric Number: 127278
Identity Card Number: 951205-07-5094
Date of Submission: 17th May 2016 (Tuesday)

Content
Contents
1.0 Objective
2.0 Hypothesis
3.0 Introduction
4.0 Materials and
Apparatus
5.0 Methods and
Procedures
6.0 Results
7.0 Discussion
8.0 Conclusion
9.0 References

Pages
3
3
3-5
6
6-8
8-9
10-15
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1.0 Objective
The objectives of carrying out this experiment are:
a To study the linked gene traits in Drosophila melanogaster by constructing the
b

inheritance cross diagram for each cross of Drosophila melanogaster.


To identify the gene order and the map distance between the genes of Drosophila
melanogaster using gene mapping.

To construct the genetic map involved in each cross of Drosophila melanogaster


through the analysis of results of the gene map distance.

2.0 Hypothesis
The hypothesis that we can made from this experiment are:
a The linkage map distance between the gene loci can be determined by calculating
b

the percentage of recombinant between two genes.


The genetic map of Drosophila melanogaster can be drawn from the results of
their gene order and linkage map distances.

3.0 Introduction
There are two laws for Mendels Law of inheritance that is the principle of segregation
and the principle of independent assortment. For the first law, the two members of a gene pair
that are the alleles segregate from each other in the formation of gametes. Half the gametes
carry one allele and the other half carries the other allele. Whereas for the second law, genes
for different traits assort independently of one another in the formation of gametes. However,
in some cases the genes are not assorted independently. This is because the genes that are
nearer or closer to each other on the same chromosome may tend to be genetically linked. In
other words, the closer the two genes are on the chromosome, the lower is the tendency for
the chance of swapping to occur between them and the higher chances they are to be
inherited together. Therefore, the tendency for the alleles that are close together on a
chromosome to be inherited together during the meiosis phase of sexual reproduction is
known as genetic linkage.

Genetic linkage was first discovered by British geneticists shortly after the Mendels
laws were discovered. The geneticists who discover genetic linkage is William Bateson, Edith
Rebecca Saunders and Reginald Punnett. After a period of time, the understanding of the
genetic linkage was enhanced by the work of Thomas Hunt Morgan as he had successfully
constructed the first gene mapping work of Drosophila melanogaster and thus providing the
important evidence for the chromosome theory of inheritance. Drosophila melanogaster has
been used as a model organism in genetic analysis and discoveries from long time ago until
now because it is small in size, having short life cycle, high reproductive rate and ease of
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culture, has only four pair of chromosomes and genetic manipulation have made it perhaps
the best understood animal genetic system.
Alfred Sturtevant, a student of Morgan's, first developed genetic maps, also known as
linkage maps. Gene mapping describes the methods used to identify the locus of a gene and
the distances between genes. Besides that, gene mapping brings lots of benefits in identifying
gene traits, the linkage map distance between the genes and the position of locus of the gene.
For example for identifying gene traits is to identify whether the gene is linked or unlinked.
From the genetic map which has been constructed, the relative position of allelic
characteristics on the Drosophila chromosome and the gene map distance between gene loci
can be shown clearly. The map distance between the gene loci can be determined by using the
formula:

Map distance =

number of offspring withrecombinant phenotype


100
total number of offspring

Recombinant occurs due to the crossing over during meiosis as the portions of
chromatids of homologous chromosome breaks and reunions at chiasmata. Recombination
frequency refers to a measure of genetic linkage and it is useful in the creation of a genetic
linkage map. Recombination frequency is the frequency with which a single chromosomal
crossover will take place between two genes during meiosis. A centimorgan (cM) or a map
unit (mu) is defined as the distance between genes for which one product of meiosis in 100 is
recombinant. Besides that both the units used to describe a recombination frequency of 1%.
In this way, we are able to measure the genetic distance between two loci based on their
recombination frequency and this is a good estimate of the real genetic distance between two
loci.
In genetics, the coefficient of coincidence (c.o.c.) is a measure of interference in the
formation of chromosomal crossovers during meiosis. It is generally the case that, if there is a
crossover at one spot on a chromosome, this decreases the likelihood of a crossover in a
nearby spot. This is called interference. The coefficient of coincidence is typically calculated
from recombination rates between three genes. For examples, if there are three genes in the
order A B C, then we can determine how closely the genes are linked by frequency of
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recombination. Knowing the recombination rate between A and B and the recombination rate
between B and C, we would naively expect the double recombination rate to be the product
of these two rates. Thus, the coefficient of coincidence is calculated by using the formula
given below:

With the value of coincidence, the value of interference is defined by the formula: coefficient
of interference equals to 1- coefficient of coincidence. The value of coefficient of coincidence
actual double recombinant frequency
and interference tell us how strongly a crossover in one of the DNA interferes with the
Coefficient of coincidence
= expected double recombinant frequency
formation of a crossover in the other region.

4.0 Materials and Apparatus


The materials and apparatus that we used in this experiment include:

Wild type male Drosophila melanogaster


Yellow body, singed bristle and white eyes female Drosophila melanogaster
Black body, vestigial wing and brown eyes female Drosophila melanogaster
Four clear vials tube with cotton plug
Small paint brushes
Sheets of A4 white paper
Ether bottle
Ether Solution
Label

5.0 Methods and Procedures


A Transferring flies into vials containing food medium
1 Cotton is placed under the bottle cover cap and few drops of ether are dropped
onto the cotton. The bottle is closed for a few seconds until the ether gas has
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filled the entire bottle.


The vials containing the flies are knocked so that they are drop on the bottom

of vial to prevent them from flying away.


The flies are then transferred into an ether bottle which has cotton fitted with a

cotton plug.
The flies will become immobilized or fainted within a few seconds. However,
the flies cannot be exposed to ether for too long as this will cause them to die

and not fainted.


Small paint brushes are then used to transfer the flies into the fresh vial

containing the food medium.


The vial is remained horizontally until the flies are awaked to prevent the flies

from sticking in the agar solution.


Finally, the leftover Drosophila melanogaster are discarded into an alcohol
concentrated.

B Method of making cross breed


1 One of the vial tube, 3 female Drosophila melanogaster with yellow body,
single bristles and white eyes (y sn w / y sn w) are crossed with 3 wild type
2

male Drosophila melanogaster (+ + + / 0).


In another vial tube, 3 female Drosophila melanogaster with black body,
vestigial wing and brown eyes (b vg bw / b vg bw) are crossed with another 3

3
4

wild type Drosophila melanogaster (+ + + / + + +).


All vials are then labelled with type of cross, genes, date and group name.
The vials are incubated at the room temperature and away from bright light to

ensure a successful result.


The parent flies are removed after one week and the F1 generation are left to

mature.
After a few days, all the flies from the culture vial are cleared from the vial

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tube and the virgin females are collected a few hours later.
During day 15, another two cross breed are carried out.
In the first cross breed, the virgin heterozygous female flies are crossed with
male flies (b vg bw / b vg bw).

While, in second cross breed, the female flies ( + + + / y sn w) are crossed

with male flies ( y sn w / 0)


10 One week later, the F1 parents are removed again and F2 generation are left to
mature.
The days, dates and types of activities are showed below:
Days
1

Dates
31 Mac 2016

Activities
Parental cross breed
y sn w/ y sn w female x + + + / 0
male
b vg bw / b vg bw x + + + / + + +

8
12
15

07 April 2016
12 April 2016
14 April 2016

male
Remove parent flies
Collect virgin females (8 a.m.)
Virgin heterozygous female x b vg bw
male
+ + + / y sn w female x y sn w/ 0

22
24

21 April 2016
24 April 2016

male
Remove F1 flies
Collecting and Counting F2 generation

C) Method of Counting the F2 generation


1 All F2 generation flies are killed by exposing them to ether.
2 Dead flies are transferred onto a sheet of white paper and observed under the
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4
5

microscope to identify their bristles characteristics.


The flies are separated according to their phenotype.
The numbers of flies are calculated and recorded as results in table form.
The inheritance cross diagram for each cross are constructed and the map
distance between genes, coefficient of coincidence, C and interference, I are
calculated. Genetic maps for each cross are then been constructed.

6.0 Results
A Cross breed between b vg bw/ b vg bw with + + + / + + +
b vg bw
b+ vg+ bw+
b vg bw
b+ vg+ bw+
b
vg Xbw
b+ vg+ bw+

37
29
7

b
vg+ bw+
16
+
b
vg
bw
13
+
b vg
bw
5
b+ vg+ bw
26
b
vg+ bw
3
+
b vg bw+
2
*Note: This experiment results were obtained from our senior because the experiment carried
out by us was unsuccessful.

B Cross breed between y sn w / y sn w with + + + / 0


y sn
w
y+ sn+ w+
y sn
w
.

The position of gene should be y w sn since the results of this genes position
produces double-crossover phenotypes which match with the results.

y
y+
y+
y
y
y+
y
y+

w
w+
w+
w
w+
w
w+
w

sn
sn+
sn
sn+
sn+
sn
sn
sn+

57
200
29
6
8
1
0
0

*Note: This experiment results were obtained from our senior because the experiment carried
out by us was unsuccessful.

7.0 Discussion
1 Draw an inheritance cross for both crosses.
A Cross breed between b vg bw/ b vg bw with + + + / + + +
Parents

: b vg bw
b vg bw

Gametes

F1 generation

F1 x

b vg bw
b vg bw

Gametes

vg

b+ vg+ bw+
b+ vg+ bw+

bw

b+ vg+ bw+
b vg bw

b+ vg+ bw+
b
vg bw

: b

vg bw

b vg bw
b vg bw

b+ vg+ bw+

b vg+ bw+

b+ vg bw

bw+

b+ vg+ bw

b vg

b vg+ bw

F2 generation:

b+ vg+ bw+

b vg bw

b+ vg bw+

b vg bw / b vg bw
b vg+ bw+/ b vg bw

b+ vg+ bw+ / b vg bw
b+ vg bw / b vg bw

b vg bw+/ b vg bw
b vg+ bw / b vg bw

b+ vg+ bw / b vg bw
b+ vg bw+ / b vg bw

B Cross breed between y w sn / y w sn with + + + / 0


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Parents

y+ w+

sn

sn+

sn

0
Gametes
w+

sn+

F1 generation
F1 x

y+

sn

y sn w
0

y+ w+ sn +
y w sn

:
y+ w+ sn+
y w sn

w sn
0

X
Gametes

sn

y+ w+ sn
y
y
F2
:

w+ sn+
w+ sn

y+ w+ sn+
y

sn+

y w sn
0

y+ w sn
y+ w sn+

y w sn / y w sn
y+ w+ sn / y w sn

y+ w+ sn+ / y w sn
y w sn+ / y w sn

y w+ sn+/ y w sn
y w+ sn / y w sn

y+ w sn / y w sn
y+ w sn+ / y w sn

y w sn / 0
y+ w+ sn / 0

y+ w+ sn+ / 0
y w sn+ / 0

y w+ sn+/ 0
y w+ sn / 0

y+ w sn / 0
y+ w sn+ / 0

generation

2 Calculate the map distance between genes.


A Cross breed between b vg bw/ b vg bw with + + + / + + +

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Black-coloured body
Brown-coloured body
Vestigial Wing
Full Wing
Brown Eyes
Red Eyes

b
vg
vg+
bw
bw+

F2 phenotype
+

NCO

b
b

vg bw
vg
bw

Observed

Total

Number
29
37

Number
66

SCO

b+ vg+
b
vg

bw
bw+

26
5

31`

SCO

b+ vg
bw
+
b
vg bw+

13
16

29

DCO

b+ vg
bw+
b
vg+ bw

2
3

Total
Notes: NCO =non crossing over, SCO = single crossing over, DCO =
double crossing over

Map distance between b and vg

10

Map distance between vg and bw =

Map distance between b and bw

131

5+ 29
131
5+ 31
131

x 100 = 25.95 cM

x 100 = 27.48 cM

= 25.95 cM + 27.48 cM = 53.43

cM

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B Cross breed between y w sn / y w sn with + + + / 0


y
y+
sn
sn+

Yellow coloured body


Brown coloured body
Singed bristle
Normal bristle
White eyes
Red eyes

w
+

F2 phenotype

Observed

Total

Number
200
57

Number
257

29
6

35

NCO

y+ w+ sn+
y w
sn

SCO

y + w+
y w

SCO

y+ w sn
y w+ sn+

8
1

DCO

y+ w sn+
y w+ sn

0
0

sn
sn+

Total
Notes: NCO =non crossing over, SCO = single crossing over, DCO =
double crossing over

Map distance between y and w

Map distance between sn and w =

29+ 6+0+0
301
8+1+ 0+0
301

x 100 = 11.63 cM

x 100 = 2.99 cM

Map distance between y and sn = 11.63 cM + 2.99 cM = 14.62 cM


3 Calculate the coefficient of coincidence, C and coefficient of interference, I.
A Cross breed between b vg bw/ b vg bw with + + + / + + +
Expected DCO

= 0. 2595 x 0.2748 = 0.0713

Observed DCO

= 5/131= 0.0382
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301

Coefficient of coincidence =

0.0382
0.0713

= 0.5358

=53.58%
Coefficient of interference = 1- 0.5358 = 0.4642
= 46.42%
Results explanation:
The positive value of interference occurs when the coefficient of coincidence is
less than 1. In such cases, the positive interference value indicates that the
occurrence of a crossover in one region of a chromosome will decreases the
probability of the second crossover to occur nearby it and thus give a lower
number of double crossovers.
B Cross breed between y w sn / y w sn with + + + / 0
Expected DCO
Observed DCO

= 0.1163 x 0.0299 = 0.0035


= 0/301 =0
0
Coefficient of coincidence = 0.0035 = 0
Coefficient of interference = 1- 0 = 1

Results explanation:
In this cross breed the coefficient of coincidence is 1. So the interference is
complete and no double crossover progeny are observed.

Map out all the genes involved in each cross.

A Cross breed between b vg bw/ b vg bw with + + + / + + +


Genetic map:
25.95 cM
b

27.48 cM
vg
53.43 cM

bw

Cross breed between y w sn / y w sn with + + + / 0


Genetic map:
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2.99cM
y

11.63 cM
w
14.62

sn
cM

8.0 Conclusion
The objective of this experiment is to study the gene mapping in Drosophila
melanogaster. Through the gene mapping, we are able to identify the gene traits in
Drosophila melanogaster, the gene order and the map distance between genes of Drosophila
melanogaster. Moreover, by calculating the coefficient of coincidence and interference, we
are able to determine how strongly a crossover in the gene of Drosophila melanogaster may
interfere with the formation of a crossover in another region.
From the results of coefficient of interference that we obtained from the cross breed
between b vg bw/ b vg bw with + + + / + + +, we are able to concluded that the positive value
of interference indicates that the occurrence of one exchange that is the crossing over
between homologous chromosomes appears will tend to reduce the likelihood of another in
its vicinity. In a simpler way of saying, the occurrence of a crossover at one spot on a
chromosome will decreases the likelihood of another crossover to be occurring in a nearby
spot.
Besides that, the results of coefficient of interference that we obtained from the cross
breed between y w sn / y w sn with + + + / 0, we are able to concluded that the positive value
of interference indicates the interference is complete and no double crossover progeny are
observed. This is because in this cross breed the coefficient of coefficient is 1.
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9.0 References
1) Genetic lingkage. Wikipedia, the free encyclopedia (17th April 2016). Retrieved from
https://en.wikipedia.org/wiki/Genetic_linkage
2) Pearson.BioCoachActivity.

Retrieved

from

http://www.phschool.com/science/biology_place/biocoach/inheritance/laws.html
3) Gene Mapping. Wikipedia, the free encyclopedia (15th March 2016). Retrieved from
https://en.wikipedia.org/wiki/Gene_mapping
4) Interference-An Introduction to Genetic Analysis_NCBI Bookshelf. AJF Griffiths - 2000.

Retrieved from http://www.ncbi.nlm.nih.gov/books/NBK22051/


5) Genetics: Ch.7 flashcards | Quizlet. Andrew Sutherland. January 2007. Retrieved from
https://quizlet.com/28047101/genetics-ch7-flash-cards/
6) Coefficients of coincidence. Retrieved from
https://en.wikipedia.org/wiki/Coefficient_of_coincidence

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