CHROMOSOMES

First observed in tumor cells (cells are actively dividing thus able to see a lot of nuclei containing
chromosomes) by Walter Fleming in 1882, 16 years after establishment of genetics.

EUKARYOTIC CHROMOSOMES

- More than one

- Linear rather than circular
- Sequestered within a nucleus
- Composed of DNA and globular proteins
(histones)
- Nucleosomes (10 nm diameter beads)
- Chromatin (30 nm diameter)
- Active genes, NOT visible – Euchromatin
- Inactive DNA, visible, located at the periphery
of nuclei – Heterochromatin

Examples:

- Human chromosomes: 16 (44 autosomes, 2

sex chromosomes)
- Fruit fly (2x: 8)
- Kingfisher (2x: 132)
- Tobacco (4x: 48)
- Adder’s tongue fern (2x: 1260)

EXTRANUCLEAR DNA: EUKARYOTES

 Mitochondria (Animals), Chloroplasts (Plants)

- Some DNA are CIRCULAR; Human cells have LINEAR
- DNA codes for 5% RNA and polypeptides (nonfunctional) required for the organelle
replication and function
 Plasmid
- Fungi and protozoa
- Example: S. Cerevisiae – contains 70 copies of a plasmid (2 μm circle) - μm (micron) one
millionth of a metre
 PROKARYOTIC CHROMOSOMES

 Consists of a single molecule of DNA in the form of a

closed loop
Exceptions:
- Agrobacterium temefaciens = 1 linear, 1 circular
- Vibrio cholerae = 2 circular
 Bacterial-circular molecule of DNA is associated with
proteins and RNA
- Located in NUCLEIOD
- Folded into loops that are 50,000 – 100,1000 base pairs

PLASMIDS

- Small, circular molecules of DNA

- Few thousand base pairs to million base pairs
- Carries information for replication, cellular traits

Types of Plasmids
(F) factors
1. Fertility (F) factors – conjunction, transfer of genes

2. Resistance (R) factors – carry genes for resistance to one or

more antimicrobial drugs, heavy metals or toxins
Transmission of genetic material
3. Bacteriocin factors – carry genes for proteinaceous toxins to the new daughter bacterium

(bacteriocin).

4. Virulence plasmids - carry genes for pathogenicity

Parts of a Chromosomes

 Telomere – The ends of the chromosome

 Centromere – The primary constriction of the
chromosome; It also divides the chromosome into
short arm (p) and a long arm (q)
 Chromatid – A single molecule of DNA

Classification of Chromosomes

- Size and position of centromere in the middle of the

chromosome
 Metacentric – centromere in the middle of the
chromosome
 Submetacentric – centromere divides the
chromosomes into 1/3 and 2/3
  Acrocentric – Centromere near the end of the chromosome

Scientist called cytogeneticists can recognize and identify many of these gross chromosomal
abnormalities by examining chromosomes through a microscope

CYTOGENETICISTS USE THREE THINGS TO TELL CHROMOSOMES APART:

1. Chromosome SIZE

2. the POSITION of the centromere

3. Characteristic BANDING PATTERNS of alternating light and dark bands (caused by staining the
chromosomes with dyes)

DETECTION

1. Classic (depicted) Karyotype

- A dye, often Giemsa (G-banding), is less frequently Quinacrine, is used to stain bands on the
chromosomes.
- Each chromosome has a characteristic banding pattern
that helps to identify them; both chromosomes in a pair
will have the same banding pattern
 Giemsa
- Specific for PHOSPHATE GROUPS of DNA
 Quinacrine
- Binds to the ADENINE-THYMINE-RICH
REGION
 Karyotypes are arranged with the short arm of the chromosomes on TOP, and the long arm on the
BOTTOM
 The short and long arms are called p and q, respectively.
 In addition, the differently stained regions and sub-regions are given NUMERICAL DESIGNATIONS
from proximal to distal on the chromosome arms.
2. Spectral Karyotyping

- Is a molecular cytogenetic technique used to simultaneously visualize all the pairs of chromosomes
in an organism in different colors
- Fluorescently labeled probes for each chromosome are
made by labeling chromosome-specific DNA with different
fluorophores.
- Due to limited number of spectrally-distinct fluorophores,
a combinatorial labeling method is used to generate
many different colors.
- Spectral differences generated by combinatorial labeling
are captured and analyzed by using an interferometer
attached to a fluorescence microscope.
- Image processing software then assigns a pseudo color to each spectrally different combination,
allowing the visualization of the individually colored chromosomes.
- The more color used, the more expensive it is

3. Digital Karyotyping or Virtual Karyotyping

- A technique used to quantify the DNA copy number on

genomic scale
- Short sequences of DNA from specific loci all over the
genome are isolated and enumerated
- Subjected with dye (Giemsa or Quinacrine)