Course Course Course L T P C

C
Code 21BTC206T Name GENETICS AND CYTOGENETICS Category PROFESSIONAL CORE 3 0 0 3

Program Outcomes (PO) Program

Specific
Outcomes
1 2 3 4 5 6 7 8 9 10 11 12

Conductinvestigations of complex
Design/development of solutions

Environment&Sustainability
The engineer and society

Individual&Team Work
Engineering Knowledge

Project Mgt. &Finance

ModernTool Usage

Life LongLearning
ProblemAnalysis

Communication
problems

Ethics

PSO-1

PSO-2

PSO-3
CLR-1: explain the concepts and experiments in the preparation of linkage map
3 3 3 2 - - - - - - - - 3 - -
CO-1: construct linkage maps from inheritance pattern of different genes

Unit-2 - Linkage and Chromosome Mapping

Chromosome structure, Chromosome organization, Giant chromosomes - polytene chromosome, Lampbrush
chromosome; Linkage - Arrangement and types of linkage; Crossing over - Frequency of recombination, Cytological
basis of crossing over - Stern’s experiment; Chromosome mapping - Mapping by two factor cross, Mapping by three
factor cross, Interference and Coincidence, Solving Problems, Combining of map segments, Preparation of linkage
map; Somatic cell hybridization - HAT selection procedure
LINKAGE AND CHROMOSOME MAPPING
 Looped domains attach to the inside of the nuclear envelope
How histones influence folding in eukaryotic DNA
 Histone structure

Nucleosome organization

Nucleosomes are the building blocks of chromatin

PowerPoint Presentation (biophysics.org)
Heterochromatin

Chromatin that remains condensed when other chromatin are less condensed, replicate late in S phase
and aren't genetically active, Barr body, telomeres, areas adjacent to centromeres
 Heterochromatin Euchromatin

Highly condensed Decondensed

Repetitive sequences Single copy sequences (genes)

Replicates later in the cell cycle Replicates early in the cell cycle

Transcriptionally OFF Transcriptionally ON

Giant chromosomes
Some cells at certain particular stage of their life cycle contain large nuclei with giant or large sized
chromosomes. Polytene and lampbrush chromosomes are examples of giant chromosomes.
Lampbrush chromosome
Lampbrush chromosome
Lampbrush chromosome
 Lampbrush chromosome

•Lampbrush chromosomes were first observed by Flemming in 1882 in sections of Salamander

oocytes and later described by Ruckert in the year 1892.
•They appeared like brushes used for cleaning lamps, hence the name lampbrush chromosome.
•They are transitory structures and can be observed during the diplotene stage of prophase I in
meiosis in the oocytes of all animal species both vertebrates and invertebrates.
•They have been described in Sepia (Mollusca), Echinaster (Echinodermata) and in several species
of insects, shark, amphibians, reptiles, birds and mammals (humans).
•Lampbrush chromosomes have also been found in spermatocytes of several species, giant nucleus
of Acetabularia and even in plants.
•Generally they are smaller in invertebrates than vertebrates. They are observed in oocytes because
oocytes are high in DNA content.
•Lampbrush chromosomes are functional for studying chromosome organization and genome
function during meiotic prophase.
•Additionally lampbrush chromosomes are widely used for construction of detail cytological maps
of individual chromosomes.
 Lampbrush chromosome

•They are of exceptionally large sizes and present in bivalent

form.
•They are formed due to the active synthesis of mRNA
molecules for future use by the egg cells, when no synthesis
of mRNA molecule is possible during the mitotic cell
division.
•Lampbrush chromosomes are clearly visible in the light
microscope they are organized into a series of chromomeres
with large chromatin symmetrical loops extending laterally.
•Each loop appears at a constant position in the chromosome
(10,000 loops per chromosome set or haploid set).
•Each loop has an axis made up of DNA unfolded from the
chromosome and is transcriptionally highly active. Wherein
several transcription units with polarized RNP-matrix coats
the DNA axis of the loop.
•The majority of the DNA, however, is not in loops but
remains highly condensed in the chromomeres on the axis and
lacks expression of genes.
Polytene Chromosome
 Polytene Chromosome

•Giant chromosomes were first time observed by E.G. Balbiani in the year 1881 in nuclei of certain secretory
cells (salivary glands) of Chironomas larvae (Diptera).
•However he could not conclude them to be chromosomes. They were conclusively reported for the first time
in insect cells (Drosophila) by Theophilus Painter of the University of Texas in the year 1933.
•Since they were discovered in the salivary glands of insects they were termed as salivary gland chromosomes.
•The name polytene chromosome was proposed by Kollar due to the occurrence of many chromonemata
(DNA) in them.
•Cells in the larval salivary gland of Drosophila, mosquito and Chironema contain chromosomes with high
DNA content.
•However they may also occur in malphigian tubules, rectum, gut, foot pads, fat bodies, ovarian nurse cells etc.
•Polyteney of giant chromosomes happens by replication of the chromosomal DNA several times without
nuclear division (endomitosis) and the resulting daughter chromatids do not separate but remain aligned side
by side.
 Polytene Chromosome

•During endomitosis the nuclear envelope does not rupture and no spindle formation takes place. The polytene
chromosomes are visible during interphase and prophase of mitosis.
•They are about 100 times thicker contain 1000 to 2000 chromosomes, than the chromosomes found in most
other cells of the organism.
•When stained and viewed under compound microscope at 40X magnification they display about 5000 bands.
In them the chromomere or the more tightly coiled regions alternate with regions where the DNA fibres are
folded loosely.
•A series of dark transverse bands alternates with clear zones of inter bands. Such individual bands can be
correlated with particular genes.
•About 85% of the DNA in polytene chromosomes is in bands and rest 15% is in inter bands. The cross
banding pattern of each polytene chromosome is a constant characteristic within a species and helps in
chromosome mapping during cytogenetic studies.
•In Drosophila melanogaster there are about 5000 bands and 5000 interbands per genome.
 Polytene Chromosome

•These chromosomes are not inert cellular objects but dynamic structures in which certain regions
become “puffed out” due to active DNA transcription at particular stages of development.
•These chromosome puffs are also termed Balbiani rings. Puffs may apprear and disappear
depending on the production of specific proteins which needs to be secreted in large amounts in the
larval saliva.
•Another peculiarity of the polytene chromosomes is that the paternal and maternal chromosomes
remain associated side by side and the phenomenon is termed somatic pairing.
•Both polyteney and polyploidy have excess DNA per nucleus, but in the later the new chromosomes
are separate from each other.
•A polytene chromosome of Drosophila salivary glands has about 100 DNA molecules which are
arranged side by side and which arise from 10 rounds of DNA replication
(210=1024). Chironimus has 16000 DNA molecules in their polytene chromosomes.
Polytene Chromosome

The structure of Drosophilla polytene

chromosome. A: mRNA; B-Chromosome puff;
C: Chromonemata; D: Dark band; E: Interband
 POLYTENE CHROMOSOME VS LAMPBRUSH CHROMOSOME

Feature Polytene Chromosomes Lampbrush Chromosomes

Thickened, giant chromosomes Long, extended chromosomes with

Structure
with many bands loops

Location Found in certain tissues of insects Found in the oocytes of vertebrates

Replicates multiple times without Replicates during the diplotene
Replication
cell division stage of meiosis
Exhibits characteristic banding
Band Patterns Loops show transcriptional activity
patterns
Involved in rapid and intense gene Primarily associated with RNA
Function
expression synthesis
Present in specialized reproductive
Cell Type Present in somatic cells
cells
Discovered in Drosophila Discovered in lampreys and
Discovery
melanogaster amphibians
Reference: Giant chromosomes - Biotechtutorials
 Comparison of giant chromosomes vs normal chromosomes

Feature Giant Chromosomes Normal Chromosomes

Very large, may be up to 1000 times

Size Typically 0.5-2 μm long
longer than normal chromosomes

Number per cell One or a few Typically tens to hundreds

Typically constricted region along
Centromere Usually terminal or subterminal
length
Present at ends to prevent
Telomeres Usually absent or not distinct
fusion/damage
Each contains small fraction of total
DNA content Contains majority of cell’s DNA
DNA
Cell types Found in some protists and insects Found in most eukaryotic cells

Pairing during cell division Remain unpaired Synapse and pair along length

Segregation during cell division Segregate intact Break and segregate in pieces
Chiasma formed to enable
Recombination Little or no chiasma formation
recombination
Gene expression may be better More opportunity for genetic
Evolutionary advantage
coordinated recombination
Reference: Giant chromosomes - Biotechtutorials
In linkage, two or more genes linked together are always inherited together in the same combination for more than
two generations
• Morgan noted that while crossing a set of characters, two genes did not follow Mendel’s
law as they did not divide as per the law.
• The likelihood of achieving a parental combination if two genes are situated on the
same chromosome was relatively higher in the subsequent generation in comparison to
the non-parental combination.
• The physical connection of genes was referred to as linkage.
• Genes are said to be linked when genes for different traits are located in similar
chromosomes and hence are tied to each other.
• It is a deviation from the Mendelian principle of independent assortment that is
appropriate to be applied to the genes that are situated on different chromosomes.
Types Of Linkages
Linkages can be classified into two types established on the absence or presence of non-
parental combinations or new combinations, namely:
Complete Linkage
A linkage is said to be complete when two or multiple characteristics are inherited and
normally surface in two or further generations in their parental or original combinations,
they are known as complete linkage. These particular genes do not generate combinations
that are non-parental. The genes that exhibit these linkages are located nearby in the same
chromosome. Examples – genes for long wings and grey body in male Drosophila
Incomplete Linkage
It is displayed by genes that generate some portion of non-parental combinations. These
genes are situated at a distance on the chromosomes which can be attributed to the
occasional or accidental deconstruction of chromosomal
 Linkage And Crossing Over
• Linkage is the tendency of genes present in the chromosome to stay intact and transfer to
the next generation whereas crossing over is the exchange of chromosomal sections to
disrupt built links and form new linkages.
• Linkages generate parental types and increases in age as the linkage increases. Crossing over
generates recombinations and it decreases with age and the occurrence of crossing over between
two genes is decreased if they are placed closely, whereas the linkage strength between two
genes increases if a chromosome is located in the vicinity.
• Linkages assist in maintaining a newly improved variety, crossing over, on the other hand, forms
as a source of variations to produce new varieties.
• Crossing over, in other terms, is the exchange of segments observed in homologous
chromosomes between non-sister chromatids and takes place during the pachytene stage of the
prophase I in the cell division process of meiosis and always takes place within linked genes.
The recombination of linked genes that crossing over produces plays a significant role in evolution.
CROSSING OVER
DOUBLE CROSSOVER CAN OCCUR IN THREE DIFFERENT WAYS