Human Genome Project

Human Genome Project is the most ambitious and exciting scientific

undertaking by human being. Human genome project is administered
by National Institute of Health and US Deptt. of Energy. In U.S.A.
work started on this project in 1990 with determination to map and
sequence the complete set of chromosomes in 15 years.

Acquiring complete knowledge of the organization, structure and

function of human genome – the master blueprint of each of us – is the
broad aim of Human Genome Project.’ With the target of three billion
base pairs in human genome to be sequenced and by 1990- 100,000 had
been analysed.

Many sequences in genome are highly repetitive and have no obvious

function. This is about 50% of total DNA. As seen above, it is not a
small job and labours and cost wise frustrating too. It is an ambitious
project.

Studies of this project will be undertaken under three steps:

(i) Mapping

(ii) Sequencing

(iii) Functional analysis

Mapping of human genetics is difficult due to following reasons:

(i) Ethical reasons

(ii) Length of breeding cycle is only 15-16 years.

(iii) Inability to carry on controlled breeding.

Regular view of the strategies used for mapping, extensive automation
for sequencing and newer techniques may help the scientists to achieve
the goal.

Donnis Keller et.al. (1987) described first RFLP map of human

genome. It identified RFLP loci with an average spacing of 10
centimorgans (cM). G. Craig Venter et.al. (1990) instead of taking
‘Human Genome Project’ strategy of sequencing chromosomal DNA
one base at a time, they isolated mRNA molecules, copied these RNA
molecules into DNA sequence tags or ESTs.

They identified over 8000 genes in period from 1990- 1992. Dr. Venter
with his 70 scientists at Institute of Genomic Research, Gaitherburg,
Maryland expects to determine sequence of 2000-3000 human genes
per week.

Physical maps of all human chromosomes were complete by 1994. EST

can be used to isolate the entire gene. With EST plenty of databases of
nucleotide sequences has been made available. It has facilitated the
construction of preliminary transcript map of human genome.

ESTs represent only expressed genes. ESTs are cDNA clones. Using
nucleotide sequence databank (Gene bank) and protein sequence
database (Protein Information Resource) ESTs can be matched with
existing sequences and assigned genes with the help of computer
programmes.

Dr. Venter further devised ‘whole genome shot gun strategy’ which
involves randomly breaking DNA into segments and cloning the
fragments into vectors. The three dimensional structures of smallest
genome Mycoplasma genitalium and Haemophilus influenzae Rd
proteins with completely unknown functions have been determined as
a part of a structural genomics project.

This effort is investigating the use of structural information in assisting

protein functional assignment. Craig Venter formed “The Institute of
Genomics Research (TIGR)” in Maryland (U.S.A.). Here several
bacteria were sequenced.

The structures of the hypothetical proteins are being used to guide

studies for knowing the protein function. With more than 25 structures
determined by this time, the results are highly encouraging to know
some level of functional understanding.

The completion of human DNA sequence in 2003 coincided with 50th

anniversary of Watson and Crick’s description of the structure of DNA.
The Human Genome Project was marked by accelerated progress.
Rough draft of human genome was completed in 2000.

In February 2001 ‘Science and Nature’ published the working draft

sequence and analysis of human genome. Scientists first generated a
physical map of human genome with clones got from each chromosome
organized into series of long coting’s.

Because most of the clones are more than 100, 0000 bp long and present
techniques can resolve only 600 – 750 bp of sequence, each sequence
is to be sequenced in fragments.

By using shot gun approach followed by assembly of entire clone by

computerized identification of overlaps, sequenced DNA was made
available in database covering of entire genome. J. Craig Venter (1997)
in Celera Corporation made use of different strategy called whole
genome shot gun sequencing which eliminated the step of assembling
a physical map of genome.

Scientists sequenced DNA fragments throughout genome at random.

Decades old estimates that human has about 100,000 genes within the
3.2 X 109bp. The genome sequence however, revealed that there are
only 20,000 to 25,000 genes. Although humans evolved relatively
recently, human genome is very old.

Few interesting features of human genome are:

(i) There are more than 3.2 billion base pairs.

(ii) Human genome is estimated to have 20,000 to 25,000 genes and to

determine the sequences of 3 billion chemical base pairs that make up
human DNA.
(iii) Within the human population are millions of single base
differences called single nucleotide polymorphisms (SNPs). Each
human differs from the next by about 1 bp in every 1,000 bp.

(iv) The human genome bears gene rich areas separated by gene poor
areas called gene deserts.

(v) About 45% of our genome is made up of transposons.

(vi) Only 2 percent of genome codes the proteins.

(vii) To improve tools for data analysis.

(viii) Use of concerned technology to sectors like industries.

(ix) Address the ethical, legal and social issues (ELSI) which may
appear due to HGP.

(x) The human genome contains 3164.7 million nucleotide bases.

Largest known human gene is dystrophin which bears 2.4 million
bases, with average gene having about 3000 bases. Functions of 50
percent of discovered genes are yet to be found.

(xi) About 99.9 percent of nucleotide bases are exactly similar in all
human beings. Previously total number of genes estimated was 80,000
to 1, 40,000 but now number is said to be 30,000.

(xii) Large amount of human genome is constituted of repeated

sequences.

(xiii) Chromosome 1 bears maximum genes (2968), with Y

chromosome having lowest (231).
 Fig. Snapshot of human
genome

Laboratories and research groups

of many countries are involved in
the research on human genome.
Data collected from all these
sources is to be used and
integrated. Two noteworthy
contributors of Human Genome
Project are Centre d’Etudes
Polymorphism Humaine (CEPH)
and Genethon (a factory of
human genetics) in France.

Some of the important techniques used in the human

genome project are as follow:
(а) DNA Sequencing are the sequencing methods for determining the
order of the nucleotide bases—adenine, guanine, cytosine, and
thymine—in a molecule of DNA. There are two common methods
namely Maxam Gilbert technique which uses chemicals to cleave DNA
into fragments at specific bases; or, most commonly, the Sanger
technique.
It is also called the di-deoxy or chain-terminating method. It uses DNA
polymerase to make new DNA chains, in the presence of di-
deoxynucleotides chain terminators to stop the chain randomly as it
grows. In both cases, the DNA fragments are separated according to
length by polyacrylamide gel electrophoresis. This enables the
sequence to be read directly from the gel.

(b) Employment of Restriction Fragment-Length Polymorphisms

(RFLP).

(c) Yeast Artificial Chromosomes (YAC) is a vector (carrier) created

and used in the laboratory to clone pieces of DNA. A YAC is
constructed from the telomeric, centromeric, and replication origin
sequences needed for replication in yeast cells. The telomere is the end
of the chromosome; the centromere is the chromosome region to which
spindle fibers attach during cell division; and the replication origin
sequences are the spots where the replication of DNA starts.

(d) Bacterial Artificial Chromosomes (BAC) is a large segment of

DNA (100,000—200,000 bp) from another species cloned into
bacteria. After the foreign DNA has been cloned into the host bacteria,
many copies of it can be made. It is a large insert cloning vector capable
of handling large segments of cloned DNA, typically around 150 kb.
BACs can be propagated in laboratory strains of Escherichia coli.
These vectors are helpful in the construction of genomic libraries for
genome scale sequencing projects.

(e) The Polymerase Chain Reaction (PCR).

(f) Electrophoresis.
Applications of Human Genome
Project

Some of the different fields where human genome project application

is used are: (a) Molecular Medicine (b) Waste Control and
Environmental Cleanup (c) Biotechnology (d) Energy Sources (e) Risk
Assessment (f) DNA Forensics (Identification).

(a) Molecular Medicine:

Genetic screening will enable rapid and specific diagnostic tests
making it possible to treat countless maladies.

DNA- based tests clarify diagnosis quickly and enable geneticists to

detect carriers within families. Genomic information can indicate the
future likelihood of some diseases. The diseases where susceptibility
may be determined include heart disease, cancer, and diabetes.

(b) Waste Control and Environmental Cleanup:

In 1994, the Microbial Genome Initiative was formulated to sequence
the genomes of bacteria useful in the areas of energy production,
environmental remediation, toxic waste reduction, and industrial
processing. Resulting from that project, six microbes that live under
extreme temperature and pressure conditions have been sequenced. By
learning the unique protein structure of these microbes, it may be
possible to use the organisms and their enzymes for such practical
purposes as waste control and environmental cleanup.
(c) Biotechnology:
Sales of biotechnology products are projected to be very high in U.S.A.
by the year 2000. The HGP has stimulated significant investment by
large corporations and promoted the development of new
biotechnology.

(d) Energy Sources:

Biotechnology will be important in improving the use of fossil-based
resources. Increased energy demands require strategies to circumvent
the many problems with today’s dominant energy technologies.
Biotechnology will help address these needs by providing a cleaner
means for the bioconversion of raw materials to refined products.
Additionally, there is the possibility of developing entirely new
biomass-based energy sources. Having the genomic sequence of the
methane-producing microorganism.

(e) Risk Assessment:

Scientists know that genetic differences cause some people to be more
susceptible than others to such agents. More work must be done to
determine the genetic basis of such variability, but this knowledge will
directly address the DOE’s long-term mission to understand the effects
of low-level exposures to radiation and other energy-related agents,
especially in terms of cancer risk.

(f) DNA Forensics (Identification):

To identify individuals, forensic scientists scan 13 DNA regions, or
loci, that vary from person to person and use the data to create a DNA
profile of that individual (sometimes called a DNA fingerprint).