APPROVAL SHEET

Complete report of Genetics and Evolution “Giant Chromosome”. Which

made by:
Name : Andi Fitriani Ridwan
Reg. Number : 081 404 152
Group : V (fifth)
Class : ICP Biology

After checked by assistant and assistant coordinator, so the report accepted.

Makassar, December 2010

Assistant Coordinator, Assistant,

Misnawaty, S.Si Misnawaty, S.Si

Lecture Responsibility,

Hartati. S,Si. M.Si

NIP: 1974 04 05 2000 03 2 002
 CHAPTER I
INTRODUCTION

A. Background
Every living creature has a characteristic and specific properties that can be
distinguished from one another. In fact, my twin brother was difference. Traits or
characteristics possessed by every living being is declining. Characteristics and
properties that declined in every living being is controlled by genes located on
chromosomes in the nucleus of cells. Chromosomes are the carriers of heredity,
chromosomes will be more easily seen if observed if used special staining techniques
(eg, acetocarmin) at the time the cell divides. At the time, chromosomes shortened so
that it becomes thicker and can suck better than the dye contained chromosomes
when at rest. Chromosome size varies between one species and other species.
We have seen that in the cell nucleus there, and in the nucleus are
chromosomes that are smooth-shaped objects such as straight or curved rod and
consists of substances that can bind dyes within the nucleus. Chromosomes contain
the genetic substance of the DNA and act as carriers of properties. Chromosome
contained within a cell is never the same size. Similarly, the amount, the number of
chromosomes in the nucleus of every living thing that is different is different, and the
number to every living thing is constant. For example, on chromosome (Drosophila
melanogaster), which amounted to four pairs of relatively larger size and so-called
giant chromosomes.
Chromosomes are divided into two, namely chromosomes called autosomes
or chromosome body and sex chromosome or sex chromosome. Fruit fly (Drosophila
melanogaster) has 8 chromosomes is 6 autosomes and 2 sex chromosomes. Giant
chromosomes in the fruit fly (Drosophila melanogaster), will appear evident in the
phase interface. This giant chromosome formed from repeated duplication of
chromosomes without the division. According to the theory that there are
chromosomes in the form of long ribbon structure and on this tape would appear
bright and dark parts alternating. Based on the description above, it is practical to
know how the "Multiple alleles."
B. Purpose
The purpose of this lab is to observe the giant chromosomes in the larval fruit fly
(Drosophila melanogaster).
C. Benefits
The benefits of this practice are:
1. Given information to students about the giant chromosomes in the larval fruit fly
(Drosophila melanogaster).
2. This report could be used as a reference the next experiment..
 CHAPTER II
PREVIEW OF LITERATURE

Thomas Hunt Morgan and his student, however, did not await formal
cytological proof of crossing over before exploiting the implication of Janssens’s
hypothesis. They reasoned that genes located close together on a chromosome would
assort with one another much more regulary (close linkage) that genes located far
apart on a chromosome. They immediately saw this as a way to located (map) the
relatively positions of genes on chromosomes and thus to produce a genetic map. The
way they used the frequencies of the various recombinant classes is very
straightforward. Consider the segregation of three genes all located on the same
chromosome. The arrangement of the gene can be determined by means of three
crossers, in each of which two genes are followed (two factor crosses)
(Watson, 2004).
The chromosomes are the packages of genes that allow exact passage of
genetic information from one generation to the next from cell to cell as tissues grow,
and from parents to child as families grow. When the cells are doing their thing brain
cells thinking and liver cells storing glucose, for example the chromosomes are so
uncoiled inside the cell nucleus that they are not visible. It is only during the process
of cell division when the chromosomes shorten by coiling up that they can be seen as
separate bodies (Jackson, 1996).
The fruit fly D. melanogaster, a small insect 3mm long, has been used for
decades to elucidate various developmental processes through its powerful genetics.
Its importance for biological research and human health was recognized by the award
of the Nobel Prize in Physiology and Medicine to E.Lewis, C. Nusslein-Volhard and
E. wieschaus in 1995. Part of the reason why several scientist use this insect as a
model for their research is historical (so much is known about it that it is easy to
handle and manipulate) and part is practical; it is a small, with a short life cycle of
about 2 weeks, and is cheap and easy to keep in large numbers. The Drosophila egg
is about half a millimeter long. It takes 1 day for the embryo to develop and hatch
into a warm like larva (Schatten, 2008).
Therefore, other features typical of mammalian and plant polytene
chromosomes, such as noncomplete union of sister chromosomes along the length of
the chromosome, lack of a clear band pattern and temporary character of the
chromoneme union in the polytene chromosome, are also peculiar to polytene
chromosomes in some dipteral species. Classic polytene chromosomes can lose some
signs of polyteny at certain ontogenetic stages. Thus, in the larva abdominal
epidermis and enocyte nuclei of Calliphora erythrocephala, at certain ontogenetic
stage, typical polytene chromosomes dissociate into oligonate fibrils that form a
meshwork in the nucleoplasm and attach only in short regions so that is not possible
to identify individual chromosome along their entire length (Jeon, 1996).
The chromosome arms were subdivided into regions that include distinct
landmarks helpful in the identification of each chromosome. The regions were
numbered in sequential order starting at the centromere and progressing towards the
telomere, for each arm. With development of higher resolution banding techniques
the regions have been divided into bands and sub bands based on the light and dark
staining patterns. Sequential numbering of regions, bands and sub bands facilitates
the identification of specific area of the chromosome that is involved in a
rearrangement (Miller, 2001).
Representation of the 23 paired chromosomes of the human male.
Chromosome a very long DNA molecule and associated proteins, that carry portions
of the hereditary information of an organism. Structure of a chromosome (Typical
metaphase chromosome), a chromosome is formed from a single DNA molecule that
contains many genes. A chromosomal DNA molecule contains three specific
nucleotide sequences which are required for replication: a DNA replication origin; a
centromere to attach the DNA to the mitotic spindle.; a telomere located at each end
of the linear chromosome. The DNA molecule is highly condensed. The human DNA
helix occupy too much space in the cell. Small proteins are responsible for packing
the DNA into units called nucleosomes. Stained chromosomes:
Chromosomes are stained with A-T (G bands) and G-C (R bands) base pair specific
dyes. When they are stained, the mitotic chromosomes have a banded structure that
unambiguously identifies each chromosome of a karyotype. Each band contains
millions of DNA nucleotide pairs which do not correspond to any functional structure
(Jorgenson, 1978).
Preparations are best examined on a compound microscope with phase
contrast optics. I prefer to start at a lower magnification (400x final magnification),
but switch to a higher magnification (630x or 1000x final magnification) for detailed
analyses. Bridges (1935) divided the euchromatic portions of the genome into 102
numbered divisions. Divisions 1 through 20 are on left arm of the X-chromosome.
Divisions 21 through 60 and 61 through 100 comprise the second and third
chromosomes, respectively, with the centromeres between divisions 40 and 41 for the
second chromosome and between divisions are 80 and 81 for the third chromosome.
The small fourth chromosome includes divisions 101 and 102. Each division is
subdivided into six lettered subdivisions (letters a through F) in the revised maps,
each polyten chromosome band is numbered (Sullivan, 2000).
Polytene chromosomes are giant chromosomes common to many dipteran
(two-winged) flies. They begin as normal chromosomes, but through repeated rounds
of DNA replication without any cell division (called endoreplication), they become
large, banded chromosomes (see figure). For unknown reasons, the centromeric
regions of the chromosomes do not endoreplicate very well. As a result, the
centromeres of all the chromosomes bundle together in a mass called the
chromocenter. Polytene chromosomes are usually found in the larvae, where it is
believed these many-replicated chromosomes allow for much faster larval growth
than if the cells remained diploid. Simply because each cell now has many copies of
each gene, it can transcribe at a much higher rate than with only two copies in diploid
cells. The polytene chromosomes at the right are from the salivary glands of the fruit
fly Drosophila melanogaster. The bands on each chromosome are like a road map,
unique to each chromosome and well defined enough to allow high resolution
mapping of each chromosome. The Drosophila genome project uses polytene
chromosomes as a framework for the map (Baudisch, 1992).
These are giant chromosomes found in the salivary gland cells of the fruitfly
Drosophila. They are many times larger than the normal chromosomes reaching a

length of 2000 and are visible even under a compound microscope. The polytene
chromosomes appear to contain five long and one short arm radiating from a central
point called chromocentre. It is formed by the fusion of centromeres of all the eight
chromosomes found in the cell. Of the 6 arms, the short arm represents the fused IV
chromosome and the longest represents the fused sex chromosomes. These arms
contain numerous chromonemata resulting from repeated replication of DNA,
without separation into daughter chromosomes. The arms show characteristic dark
bands and light bands. The dark bands are euchromatic regions. Some of the dark
bands temporarily swell up and form enlargements called chromosomal puffs or
Balbiani rings. These regions contain actively transcribing DNA involved in the
synthesis of RNA types (Werle, 2004).
To increase cell volume, some specialized cells undergo repeated rounds of
DNA replication without cell division (endomitosis), forming a giant polytene
chromosome. Polytene chromosomes form when multiple rounds of replication
produce many sister chromatids that remain synapsed together. Polytene
chromosomes were originally observed in the larval salivary glands of Chironomus
midges by Balbiani in 1881. In addition to increasing the volume of the cell's nuclei
and causing cell expansion, polytene cells may also have a metabolic advantage as
multiple copies of genes permits a high level of gene expression. In Drosophila
melanogaster, for example, the chromosomes of the larval salivary glands undergo
many rounds of endoreplication, to produce large amounts of glue before pupation.
Polytene chromosomes have characteristic light and dark banding patterns which can
be used to identify chromosomal rearrangements and deletions. Dark banding
frequently corresponds to inactive chromatin, while light banding is usually found at
areas with higher transcriptional activity. The banding patterns of the polytene
chromosomes of Drosophila melanogaster were sketched in 1935 by Calvin B.
Bridges, in such detail that his maps are still widely used today. The banding patterns
of the chromosomes are especially helpful in research, as they provide an excellent
visualization of transcriptionally active chromatin and general chromatin structure
(Balbiani,1881).
 CHAPTER III
EXPERIMENT METHOD

A. Time and Place

Day / date : Friday, November 26th 2010
Time : At 13.20 am until 15.20 pm
Place of experiment : At the 2nd floor of Biology laboratory, the east part
Mathematic and Science Faculty, Makassar State
University.
B. Tools and Materials
1. Tools
a. Light microscope
b. Stereo microscope
c. Petri dish
d. Object glass
e. Deck glass
f. Tweezers
g. Pipette drops
h. Pin
2. Materials
a. Larva of Drosophila melanogaster
b. NaCl physiologies
c. Alcohol
d. Acetate acid
e. Acetocarmin
f. Cotton
C. Work Procedure
1. Prepared larval fruit fly (Drosophila melanogaster).
2. Soaked larvae in petri dishes which sided with physiological NaCl during 5-
10 minutes.
3. Took the petri dish then put under the stereo microscope and cut the anterior
with used the pin.
4. After that took the anterior part of larvae and took on an object glass.
5. By used a pipette drops, dropped alcohol and acetate acid to larva that has put
at object glass.
6. Then closed and pushed with deck glass.
7. Dropped with acetocarmin object glass that has closed by deck glass.
8. Deleted the excess of acetocarmin with used cotton.
9. Observed under the light microscope.
 CHAPTER IV
OBSERVATION RESULT AND DISCUSSION

A. Observation Result

Enlargement 10 X 40 Notes

1. Band
(dark part)
2. Interband
(light part)
3. Chromos
ome arm
4. Centrome
re

B. Discussion
Based on the observations that have been done, namely observation of the
giant chromosome in the larval fruit fly (Drosophila melanogaster), under the
microscope looks a giant chromosome, but its parts are not clearly visible, is caused
by a tool (microscope) used was not adequate to see the chromosomes, except that we
used larval instars did not know how many, in observation of chromosomes is needed
larval 3rd instars. Giant chromosome is located in the anterior part of the fruit fly
larvae, the chromosomes can be observed by soaking in physiological NaCl and then
member of dye that is acetocarmin so that chromosomes can be more clearly
observed under the microscope, but this can not be seen through observation by using
a microscope, considering the equipment used less than the maximum.
 According to Anonymous (2010), parts of chromosomes chromocenter polytene
namely, interband, bands, and puffs. Chromocenter is part of the chromosome
polytene as a merging of the long arm of chromosome five. Interband is the
chromosome that looks bright, because the threads of chromatin are not being rolled
up in a state but decomposes. Section is located between two bands and contains
genes that do not encode anything to transcribe. The band is part of chromosomes that
appear dark, because in part it is being rolled chromatin thread. Dark colors also are
rich is because the DNA and the genetic combination of certain genes. Puff is a
diffusion area of 5-10 bands to form a bubble structure, consisting of solid DNA that
is in the stage of transcription to produce mRNA.
According to the theory (Suryo 2007), these giant chromosomes can reach sizes
of approximately 100 times the length of chromosome body adult flies or about 500
microns. Once flattened, the length of this chromosome can reach 1180-2000 μ, the
parts consisting of the centromere of chromosomes, chromosome arms, membrane,
matrix and chromonema. Centromere is a constriction region around mid
chromosome in there kinetokor centromere. Kinetokor is the chromosome which is
where the threads of spindle attachment during division and is a place of attachment
of the core chromosome arm. Arm is the main body of the chromosome.
Chromosome arm wrapped by a membrane. Next to the membrane there is a matrix
of clear fluid that fills the entire arm. In the matrix submerged fine threads twisting
around called chromonema. Chromosome arm contains passages such as satellites
which are rounded at the ends of chromosomes and this section are not always owned
by each chromosome.
 CHAPTER V
CONCLUSION AND SUGGESTION

A. Conclusion
After done practical work, and according to our purpose, so we can conclude
that giant chromosomes found in the anterior part of larval fruit fly (Drosophila
melanogaster). Giant chromosomes of Drosophila melanogaster consists of the arms
and head chromosomes (centromere). Centromere is a constriction region around mid
chromosome. At centromere there are kinetokor and chromosome arms is the main
body of the chromosome.
B. Suggestion
1. For laboratory, better tools and materials that have damage must be substituting in
order that practical work process fluently.
2. For assistant, increase of communication with student.
3. For student, better active done practical work in Laboratory and save the clean of
Laboratory.
 BIBLIOGRAPHY

Balbiani E. G. 1881. Polytene chromosome.

http://en.wikipedia.org/w/index.php?title
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Baudisch, 1992. Polytene chromosome. http://Templates/SedatLab_Template2.dwt.

Accessed on November 30th 2010 in Makassar.

Jackson, John. F. 1996. Genetics and You. New Jersey: Humana Press Inc.

Jeon, Kwang. W. 1996. Cell Biology. California: Academic Press.

Jorgenson, J. H. 1978. Human chromosome. /Human Chromosomes_files/urchin.js.

Accessed on November 30th 2010 in Makassar.

Miller, Orlando. J. 2001. Human Chromosomes. New York: Springer Verlag.

Schatten, Gerald. P. 2008. Developmental Biology. USA: Academic Press.

Sullivan, William. 2000. Drosophila Protocols. USA: Cold Spring Harbor

Laboratory Press.

Watson, James. D. 2004. Molecular Biology of the Gene. New Delhi: Pearson
Education.

Werle, S. F, 2004. Giant chromosome. http://www.tutorvista.com/content/biology/

biology-iii/chromosomes/giant-chromosomes.php. Accessed on November
30th 2010 in Makassar.