Applied Genetic - Medical Genetic - DNA Fingerprinting & Forensic Analysis - PCR, RFLP, Electrophoresis - Human Genome Project

1. MEDICALS, DNA FINGERPRINTS & FORENSICS,
AND HUMAN GENOME PROJECT
APPLIED GENETIC:
2018
Department of Biology Education
Postgraduate Program
Palangkaraya University
CHAIDIR ADAM
ACD217004

2. BIOLOGY
MOLECULAR
Interdisciplinary Science
Biochemistry
Cell Biology
Genetics
Includes
study
MOLECULAR
biological activities on
level
has a role
Medicals, DNA
Fingerprints & Forensics,
and Human Genome
Project
INTRODUCTION

3. MEDICAL GENETICS
The progress of medical genetics has made it possible to clarify the
pathogenesis and clinical condition of monogenic (single-gene)
diseases based on the identification of the responsible genes.
• The body as machine
• Disease: The machine is broken
• Medicine: Fix the machine
• Focus is on disease
Traditional View of Disease

4. Genetical View of Disease
• Why this disease ?
• Why this person ?
• Why now ?

5. There are four types of Genetic Disease
1. Single Gene: Disorders caused by abnormality or mutation in the
sequence of one gene
2. Multifactorial: caused by a combination of environmental as well as
mutations in multiple genes
3. Chromosomal: Abnormalities in chromosome structure such as missing or
extra copies
4. Mitochondrial: caused by a mutation in the non chromosomal DNA of the
mitochondria.

6. Family History and Pedigree Analysis
Cytogenetic Studies
Fluorescence in Situ Hybridization (FISH)
DNA Analysis
Biochemical Analysis
MEDICAL GENETICS Diagnostic Techniques

7. MEDICAL GENETICS Diagnostic Techniques
Clinicians
Patient
Medical History
Parent, Siblings,
and offspring
Pertinent
Informations
Age
Sex
Ethnicity
General Health Status
Major Illnesses
Cause of Death
ask
to
obtain
FAMILY HISTORY
Pedigree
analysis
to identify
mode of inheritance
for a disease process

8. PEDIGREE
ANALYSIS
A pedigree is a diagram
representing the familial
relationships among
relatives. It can be used
to analyze Mendelian
inheritance of certain
traits
MEDICAL GENETICS Diagnostic Techniques

9. Huntington’s Disease Pedigree
Because this disease is due to a dominant mutation, all affected individuals have the
heterozygous genotype HD hd, whereas nonaffected people have the homozygous
normal genotype hd hd

10. Huntington’s
Disease

11. Albinism Pedigree
An example of a
pedigree for a
homozygous
recessive allele
is albinism

12. AA Aa aa

13. Cytogenetic
Studies 1.Cytogenetics is the study of chromosomes
utilizing light microscopy.
2.Chromosomal analysis is done by growing
human cells in tissue culture, chemically
inhibiting mitosis, staining, observing,
photographing, sorting, and counting the
chromosomes;
MEDICAL GENETICS Diagnostic Techniques

14. Cytogenetic Studies
1. Samples can be obtained from
peripheral blood, amniotic fluid,
trophoblastic cells from the
chorionic villus, bone marrow,
and cultured fibroblasts (usually
obtained from a skin biopsy);
2. In a karyotype, the
chromosomes are rearranged
systematically in pairs, from
longest to shortest, and
numbered from 1 (the longest)
through 22 to represent the
autosomes;
Prevents formation
of the spindle,
arrest cell division
during metaphase
Mitogen

15. 1. In a karyotype, the chromosomes are
rearranged systematically in pairs,
from longest to shortest, and
numbered from 1 (the longest)
through 22 to represent the
autosomes;
2. Chromosome painting, as shown in
Figure 2-4, helps to identify pairs of
homologous chromosomes. The
different colors are “painted” on each
chromosome by hybridization with
DNA strands labeled with different
fluorescent dyes.

16. Another karyotype is
shown in Figure 2-5
with chromosome
banding. These
chromosomes have
been treated with
Giemsa stain, which
causes chromosomes
to exhibit transverse
bands (G-bands) that
are specific for each
pair of homologs.
These bands allow
smaller segments of
each chromosome
arm to be identified. In
addition to allowing the
identification of
autosomes and sex
chromosomes,
chromosomal
abnormalities can be
identified through this

17. Fluoroscence In Situ Hybridization

20. DNA Analysis
1.Molecular genetics involves understanding the expression of genes
by studying DNA sequences of chromosomes.
2. Once a particular gene is shown to be defective in a given disease, the
nature of the mutation can be elucidated by sequencing the nucleotides
and comparing with that of a normal allele.
3. Molecular testing is available for more than 1000 hereditary conditions
and has had a significant impact on the diagnosis of Mendelian disorders.
4. Similar to the use of specific probes in a FISH analysis of chromosomal
abnormalities, probes are used to identify specific genes that may be
mutated in a certain hereditary disease.

21. DNA Analysis

22. DNA Analysis

23. DNA Analysis

24. Biochemical Analysis
The primary goal of biochemical testing is to determine whether certain proteins are
present or absent as well as to identify their characteristics and effectiveness in vitro.
This kind of analysis is used to look for enzymatic defects, as these important
catalysts are made of protein.

25. Of Phenylketonuria (PKU)
Biochemical Analysis

26. phenylketonuria (PKU) is an inherited disorder
caused by the absence of or a defect in the enzyme
phenylalanine hydroxylase (PAH). In the absence of
PAH, the amino acid phenylalanine accumulates and
can lead to severe mental
retardation.
Of Phenylketonuria (PKU)
Biochemical Analysis

27. Biochemical Test: PKU
Phenylketonuria (PKU)
- Inherited metablic disorder
- If untreated, leads to mental retardation,
seizures
- Affects 1 in 20,000 newborns
Missing Enzyme:
- Phenylalanine hydroxylase (PAH)
Biochemical Test:
Measure amount of Phenylalanine in Baby’s
Blood
Of Phenylketonuria (PKU)
Biochemical Analysis

28. DNA FINGERPRINT & FORENSIC
• Every individual carries a unique set of genes
• Chemical structure of DNA is same, but the order of the base pairs differs
• Every cell contains a complete set of DNA that identifies the
organism as a whole
• Only one tenth of 1% of DNA differs from person to person

29. • DNA fingerprinting is a way of telling individuals of the same
species apart
• DNA sequences are variable and can therefore be used as
identifying characteristics.
• DNA fingerprinting has advantages over other sources of evidence
(fingerprints, blood type, etc.):
• Highly accurate.
• Can be gathered from trace crime scene evidence.
DNA FINGERPRINT & FORENSIC

30. •Two Main Types of Forensic DNA Testing
•RFLP (restriction fragment length polymorphism)
• Requires larger amounts of DNA
• DNA cannot be degraded
•PCR (polymerase chain reaction)
• Less DNA and DNA can be partially degraded
• Extremely sensitive to contaminating DNA
DNA FINGERPRINT & FORENSIC

31. 1. Variable Number Tandem Repeat
(VNTR): sequences that are
repeated multiple times and the
number of repeats varies from
person to person.
2. VNTRs usually occur in introns
3. VNTRs can be amplified by PCR
and run on agarose gels to
produce unique DNA fingerprints
DNA FINGERPRINT & FORENSIC

32. DNA FINGERPRINT & FORENSIC

33. DNA FINGERPRINT & FORENSIC

34. DNA FINGERPRINT & FORENSIC

35. VISUALIZE GENE
USING GEL
ELECTROPHORESI
S METHOD
DNA FINGERPRINT & FORENSIC

36. STAIN THE
RESULT
1. Ethidium bromide
is a powerful
mutagen and is
moderately toxic
2. Be safe!!! Put
on masks and
gloves
DNA FINGERPRINT & FORENSIC

37. DNA FINGERPRINT & FORENSIC

39. DNA FINGERPRINT & FORENSIC

40. Alec Jeffreys and the Pitchfork murder case:
the origins of DNA profiling
British geneticist Alec Jeffreys began
working in 1977 on a technique that could
identify individuals through samples of
their DNA. In 1984, he and colleagues
devised a way to use a newly discovered
property of DNA, isolated areas of great
variability between individuals called
restriction fragment length
polymorphisms (RFLP), for forensic
identification—the original DNA
fingerprint.
DNA FINGERPRINT & FORENSIC

41. A fifteen year old school girl, Lynda Mann was abducted in Narbourough,
England. The next day, her body was discovered raped and
murdered. Three years later, another young woman met the same fate
near Lynda’s resting place. Richard Buckland was arrested and
confessed to the second murder only. An untested technique was applied;
‘genetic fingerprinting’ through DNA analysis. Surprisingly, there was no
match in either murder, so the test was repeated. Ultimately, Buckland was
proven innocent. As to why he confessed, he claimed he had been
pressured by police.
5,500 men from the local area were then tested. Colin Pitchfork
persuaded a friend to test in his place, but when he bragged about fooling
the investigators, he was overheard and reported. His genetic profile
matched the semen samples from both girls, and in 1987 he became the
first murderer convicted by DNA.
DNA FINGERPRINT & FORENSIC

42. HUMAN GENOME PROJECT
Goals:
1. Identify all the approximate 30,000 genes in human DNA
2. Determine the sequences of the 3 billion chemical base pairs that make
up human DNA
3. Store this information in databases
4. Improve tools for data analysis
5. Transfer related technologies to the private sector
6. Address the ethical, legal, and social issues (ELSI) that may arise from
the project

43. HUMAN GENOME PROJECT

44. HUMAN GENOME PROJECT

45. Next Generation
Sequencer
HiSeq 2500
System
HUMAN GENOME PROJECT

46. Organism Genome Size (Bases) Estimated Genes
Human (Homo sapiens) 3 billion 30,000
Laboratory mouse (M. musculus) 2.6 billion 30,000
Mustard weed (A. thaliana) 100 million 25,000
Roundworm (C. elegans) 97 million 19,000
Fruit fly (D. melanogaster) 137 million 13,000
Yeast (S. cerevisiae) 12.1 million 6,000
Bacterium (E. coli) 4.6 million 3,200
Human immunodeficiency virus (HIV) 9700 9
HUMAN GENOME PROJECT

47. HUMAN GENOME PROJECT

48. HUMAN GENOME PROJECT

49. HUMAN GENOME PROJECT

50. HUMAN GENOME PROJECT

51. HUMAN GENOME PROJECT

52. HUMAN GENOME PROJECT

53. 2018
Department of Biology Education
Postgraduate Program
Palangkaraya University
THANK YOU