•The complete genetic material of

an entire organism
is known as its genome.
 In 1986, scientists proposed a
project to make a genetic map or
catalogue, of a prototypical human,
including the chromosomal location
of all human genes and the
complete DNA sequence of the
genome
Many scientists and
physicians think that many
medical and other benefits
could flow from knowing
the location and sequence
of all the genes.
Such knowledge would
facilitate locating genes that
are associated with diseases
and disease susceptibility. It
will also make possible the
development of drugs that
are much more specifically
tailored to block particular
molecules. This effort has
become known as the
Human Genome Project.
 The Human Genome Project was
launched in the fall of 1989, and
James Watson, co-discoverer of
the double helical structure of
DNA, was appointed as the first
director.
 Watson stated his belief that the
human genome project would tell
us what it means to be human
The main goal of The Human Genome Project
was to read, letter by letter, the three billion
bases of human DNA. Before starting to
sequence the human genome, scientists built
maps of the chromosomes and developed and
refined techniques for analyzing DNA. With the
tools in place, project scientists began large-
scale DNA sequencing in 1999. In just one
year, they had amassed sequence data
covering more than 80 percent of the genome.
 The human genome is a massive
text. If the three billion letters (or
bases) of the genome were printed in
telephone books, they would require
a stack of books nearly as tall as the
Washington monument
 To accurately determine the sequence of
every base in the genome, scientists
needed to read the three billion bases not
just once, but at least six to ten times.

 Individual sequencing reactions could only

reveal the order of a few hundred bases of
DNA at a time - amounting to a fraction of
a page. This meant that to place in order
all of the DNA bases, it was necessary to
produce many thousands of overlapping
segments of DNA sequence.
 Mapping
 To begin the project, researchers built maps of
the human genome. They identified thousands
of DNA sequence landmarks that helped them
navigate across the chromosomes.
 Developing genome maps was necessary
preparation for DNA sequencing. These same
maps also served to orient geneticists who were
hunting for disease genes.
 During the Human Genome Project,
every base pair of DNA was
sequenced an average of nine times.
Some stretches of DNA were easy to
read and needed to be sequenced
little less often, while other stretches
were more difficult to read and had to
be sequenced more often.
Because the amount of
DNA in even one
chromosome is
enormous, it is not
practical to work with
the whole length of a
chromosome in
determining sequenses
The maximum size of pieces
that can be sequenced is
currently about 500-700 base
long. The chromosomes are
therefore separated and each
is cut into overlapping pieces
with restriction enzymes.
Each piece is inserted
into a plasmid which
enters bacterium. The
bacteria then divide
repeatedly and make
large quantities of one
piece at a time.
The nucleotide
sequence of each of
the pieces can then
be determined using
the establshed
method called
( di-deoxy method)
based on DNA
The DNA is used as a
template for synthesis
of new DNA strands in
a test tube.
The overall result
is the production
of a series of
smaller pieces,
each piece one
nucleotide longer
than the next
Each of the small
pieces is then
separated by
electrophoresis . The
pieces are made visible
with a fluorescent dye,
a different color is
used for each of the
four nucleotides
Fluorescent dyes make
all the pieces visible that
end with that nucleotide.
Mistakes can occur in
either copying or
sequencing, and repeating
the process does not
always give the same
answer, so the technique
must be repeated several
times in different
laboratories
After the sequence of
each piece has been
determined, the pieces
must be arranged in
their original order to get
the overall sequence
The sequence of bases in the DNA
fragment can thus be read from the
gel: the base found at the end of the
shortest piece is first ( traveled
farthest in the gel), followed by the
base found at the end of the next
longer piece ( traveled the second
farthest in the gel), and so forth.
 As each DNA fragment reaches the end of the
gel, a laser excites its fluorescent dye. A
camera detects the color of the emitted light
and passes that information to a computer.
One by one, the machine records the colors
of the DNA fragments that pass through the
gel.
 A single sequencing reaction can reveal the
order of several hundred DNA bases.

 Computer program integrates the data
from individual sequencing reactions. It
can spot where DNA fragments overlap
and order them as they originally were
on the chromosome.

 Many overlapping sequences reads are

needed to generate the uninterrupted
sequence of the original stretch of DNA.
 Whenever a stretch of DNA that spanned
2,000 or more bases was assembled, it was
placed into public databases within 24 hours.
Anyone with access to the Internet could see
and analyze the sequence data.
 After sequencing the 3 billion letters in the
human genome an average of nine times, the
Human Genome Project had released DNA
sequence for 99 percent of the genome. This
finished sequence was 99.99 percent
accurate. The project had completed all of its
goals ahead of schedule and under budget.
 Completion of the draft sequence
supported some previously
established hypotheses, but also
produced some surprises. Some key
results are:
 In February 2001 two groups
simultaneously announced completion
of draft of the sequence of the human
genome.
 Some key results are:
 About 95% of the human genome
represents noncoding DNA, a large
proportion of which is composed of
repetitive sequences.
 Less than 5% of the human genome
is composed of genes, sequences
that code for RNAs or proteins.

 It has been known for a while that

the complexity of organism does not
correlate with the size of its genome.
Much of the excess size is due to
these non-coding, repeat sequences.
 Detailed knowledge of these
sequences is opening up a new
resource for studying evolution.
 These sequences can be linked to
living fossils, carried within each of
us.
 They are already used in population
genetics studies examining the
migration of human populations.
 The actual number of genes is
smaller than previously estimated.
 In humans it is difficult to predict
which sequences represent genes.

 There are estimated 20,000–25,000
human protein-coding genes.
 The estimate of the number of
human genes has been repeatedly
revised down from initial predictions
of 100,000 or more as genome
sequence quality and gene finding
methods have improved, and could
continue to drop further.
 Surprisingly, the number of human genes seems
to be less than that of many much simpler
organisms, such as the roundworm and the
fruit fly. However, human cells make extensive
use of alternative splicing to produce several
different proteins from a single gene, and the
human proteome is thought to be much larger
than those of the abovementioned organisms.
Besides, most human genes have multiple exons,
and human introns are frequently much longer
than the flanking exons.
 A very high percentage of our genes
are not unique to humans but are
closely similar to comparable genes
from other species.
 In fact, only 1% of human genes
have no sequence similarity to any
other organism
 Mutation rates differ in different
parts of the genome. They are also
higher in males than in females,
although the reason for such a
difference is not known
 Within each gene there is an average
15 sites at which different individuals
carry a different nucleotide on each
chromosome in a pair, or at which
the same individual may have a
different nucleotide on each
nucleotide in a pair.
 These variations, called single
nucleotide polymorphisms are
greatly expanding how many alleles
are possible for different genes.
 Some of these polymorphisms are
associates with disease; most are
not, but are instead associated with
small changes in protein function or
regulation.
 Knowledge of such small scale variations
continues to challenge our concepts of
terms such as ‘heterozygous’, ‘dominant’
and ‘recessive’ and ‘allele’

 It also makes it clear that there is no such

thing as the human genome sequence. The
genome sequence of within each individual
is unique.
 This lower estimate came as a shock
to many scientists because counting
genes was viewed as a way of
quantifying genetic complexity. With
about 30,000, the human gene count
would be only one-third greater than
that of the simple roundworm C.
elegans, which has about 20,000
genes
 It could be years before a truly
reliable gene count can be assessed.
The reason for so much uncertainty is
that predictions are derived from
different computational methods and
gene-finding programs. Some
programs detect genes by looking for
distinct patterns that define where a
gene begins and ends ("ab initio"
gene finding).
 Our genes are similar to 46% of the
genes in yeast.
 More than 200 human genes and their
protein products have been found to
have significant similarity to those in
bacteria.
 These genes are not found in
intermediate organisms such as fruitflies
and one school of thoughts suggests
that these genes jumped from bacteria
to humans and vice versa.