Indian Journal of
Biotechnology
|
VOLUME 1 |
NUMBER 1 |
JANUARY 2002 |
CONTENTS
Editorial |
7 |
Papers |
|
|
Developments
in biotechnology: an overview |
9 |
|
Rintu Banerjee, Gargi Mukherjee, Ashok Pandey & A Sabu |
|
|
Some aspects
of biotechnology relevant in the national context |
17 |
|
H C Chaturvedi & Savita Agnihotri |
|
|
Biotechnology
input in fish breeding |
29 |
|
Samir Bhattacharya , Subrata Dasgupta, Malabika Datta
& Dipanjan Basu |
|
|
Heralding
the dawn of cultured adult stem cell transplantation |
39 |
Geeta Kashyap Vemuganti & D
Balasubramanian
|
|
|
Therapeutic
monoclonal antibodies: attractions and challenges |
50 |
|
|
|
|
|
|
|
Apoptosis:
an overview |
58 |
|
|
|
|
Virus
resistant transgenic plants for environmentally safe management of viral
diseases |
73 |
|
|
|
|
Electrostatic
interactions, phase separation behaviour and partitioning of proteins in
polyelectrolyte based aqueous two-phase systems |
87 |
|
|
|
|
96 |
|
|
|
|
|
101 |
|
Sudeshna Adak & Biplav
Srivastava
|
|
|
117 |
|
|
|
|
|
121 |
|
|
|
|
|
|
|
|
126 |
Indian Journal of Biotechnology
Developments in Biotechnology: An
Overview
Rintu Banerjee, Gargi Mukherjee, Ashok Pandey and A Sabu
Human quest
for improving the natural capabilities of microorganisms, making them capable
of novel processes, and discovering microorganisms with new capabilities has
led to tremendous development, today referred to as modern biotechnological
advances. This has led to the development of recombinant DNA technology, which
allows modifying microbes and other organisms to create in them highly
valuable, novel and naturally non-existent capabilities. R & D in areas
such as stem cell, human genome, transgenic crops, etc. has made untreatable
diseases to understand and control. However, research in many areas such as
cloning, stem cell, etc. has also posed serious concern on ethical and societal
issues, which if not taken proper measures, could be eventually very dangerous.
This paper traces the significant biotechnology developments and their impact
on human life with a futuristic approach.
Indian Journal of Biotechnology
Some Aspects of Biotechnology
Relevant in the National Context
H C Chaturvedi and Savita Agnihotri
Biotechnology encompasses all such plant processes, which may have practical or commercial application, albeit only temporarily a distinction may be made between fundamental and applied biotechnological research. But in all cases, priorities must be drawn for biotechnological research in the context of a developing nation, like, India with practicable objectives either realizable immediately or in the near future for economic growth of the country as well as protection and improvement of human health, putting the promise research on the back burner. Some of the aspects of Biotechnology, which may be brought to fruition with the already existing technologies and resources having down-to-earth objectives, are identified here. 1. Boosting citrus industry: In the face of the rich heritage of citrus germplasm, vast orchard area and suitable agroclimates for commercially important Citrus species, the Citrus Industry can be boosted with the application of a recent breakthrough of meristem culture if not some other techniques also. 2. Micropropagation of forest trees for reforestation: It is estimated that the green cover in the country is reduced to more than 55%, resulting in change of agroclimates, loss of biodiversity and many other related evil effects besides, shortage of wood. Micropropagation of forest trees, which is the only process to produce requisite number of plants, at a rapid rate, in the shortest possible period of time. 3. Standardization of herbal drugs: There exists a world herbal market worth $ 50 billion, in which Indian share is hardly 0.1% despite India being the home of herbal traditional drugs of Ayurveda. One of the main reasons for this is non-standard herbal formulations and gross adulteration of scientifically unidentified plant materials, in which context biotechnology can provide solutions. 4. Germplasm preservation for conservation of phytodiversity: Conservation of biodiversity, precisely phytodiversity for the survival of the human race is imperative. And the role of in vitro strategies for germplasm preservation and establishment of Gene Banks or Germplasm Repositories cannot be overemphasized. 5. Bioremediation and purification of sewage: The huge quantity of sewage is discharged everyday in the rivers, which constitute a major source for drinking water and poses a threat to the human health. As practiced by advanced countries, remediation of sewage by efficient biotechnological processes using bacteria and a few algae should be done and the purified sewage can be used for irrigation purpose, while the remaining quantity may be discharged in rivers minimising the health risk. 6. Production of bioethanol fuel: In view of the acute shortage of petrol, production of bioethanol to be used as fuel in automobiles is not only unavoidable, but also feasible by utilizing sugarcane as a raw material; sufficient quantity of sugarcane crop can be produced through well-established procedure for its micropropagation.
Indian Journal of Biotechnology
Biotechnology Input in Fish Breeding
Samir Bhattacharya, Subrata Dasgupta, Malabika
Datta and Dipanjan Basu
Pisciculture, to increase the fish production, has a crucial limitation.
Fish cultured in land-locked water bodies usually do not breed without the
hormonal induction. Technology of fish breeding suffered for a long time due to
the lack of suitable commercial product to induce the spawning of economically
important fish. Hypophysation technique, where pituitary extract was used to
induce the breeding, faced more failures than success besides the limitation of
the source. After the discovery of a brain peptide, gonadotropin releasing
hormone (GnRH), which binds to the membrane receptor of pituitary gonadotroph
cell and releases gonadotropic hormone (GTH), the situation has changed
dramatically. Final maturation and release of germ cells, spermatozoa and
oocyte, to the water for fertilization, depends on an acute surge of GTH which
is about 10 times greater than the normal circulatory level of GTH. Injection
of GnRH causes this acute GTH surge. GnRH is a decapeptide but its gene encodes
92 amino acid containing large molecule. Extensive post-translational
processing is necessary to secrete GnRH from the neural cells. For this reason
recombinant DNA technology could not be employed for the production of GnRH.
Instead, chemical synthesis of this decapeptide is easier and cheaper. Numbers
of fish GnRH structure have been elucidated and depending on salmon GnRH
peptide sequence, a chemical analogue has now been marketed under the the name
of “Ovaprim”. There is a strong research background to understand GnRH
mechanism of action and signal transduction pathway involved in GnRH mediated
GTH function on germ cell maturation and release. These scientific
investigations have contributed significantly in designing the superactive GnRH
analogues. The only GnRH available in India is from the brain of a freshwater
murrel, Channa punctatus. Combination
of two murrel GnRH variants, GnRH I and GnRH II, produces far more superior
effects than “Ovaprim”. Biotechnology input in fish breeding is no doubt highly
appreciable as it provides the cultivators almost a riskless method. However,
research in different laboratories is still in progress to have a more potent
molecule with the possible addition of some metabolic hormones. Pisciculture is
now a booming industry all over the world, which imposes a larger demand for
“Ovaprim” like products. Murrel GnRH, therefore is expected to be a highly
competitive product in the global market.
Indian Journal of Biotechnology
Heralding the Dawn of Cultured Adult
Stem Cell Transplantation
Geeta Kashyap Vemuganti and D Balasubramanian
Over the last few years, research on stem cells has received
much public attention both for its extraordinary potential and for the
associated social, legal and public issues. One of the fundamental questions in
developmental biology is: how does a single cell-the fertilized cell give rise
to a complex, multicellular organism and what are the factors involved during
this remarkable development? With that began the quest to define stem cells,
identify and isolate the embryonic, fetal and adult stem cells. Then came the
question of whether these stem cells derived from various sources can be used
to determine the pharmacokinetics of drugs and toxins; and ultimately be used
to replace cells in diseased tissues. These questions were answered by series
of experiments on animals followed by clinical trials using cultured adult stem
cells, since the latter evoke the least debate on the ethical and legal fronts.
Apart from the clinical use of hematopoietic stem cells, the other adult stem
cells that have been tested and put to clinical use include: myoblasts for cardiac
and skeletal muscle loss or degeneration; cultured beta cells for diabetes
mellitus, and cultured limbal epithelial cells for reconstruction of ocular
surface in limbal stem cell deficiency. This review briefly describes the
concept of stem cells, the regulatory mechanisms that govern them, with special
emphasis on stem cells in epithelial tissues and the technique of culturing
limbal stem cells that has been developed at our institute.
Indian Journal of Biotechnology
Therapeutic Monoclonal Antibodies:
Attractions and Challenges
Navin Khanna
Due to the
technological advances made during the past decade, therapeutic monoclonal
antibodies (MAbs) now represent an important and growing class of
bio-therapeutics. Sales of MAbs are increasing rapidly from virtually nil in
1996 to potentially over $ 1 billion in 2001. With the potential new targets
resulting from genomics and with methods now in place to make fully human
antibodies, the potential of antibodies as valuable therapeutics in oncology,
inflammation and cardiovascular disease is being fully realised. There are 235
MAbs under different stages of development. More than 100 clinical trials are in
progress, 14 of them in phase III (mostly for cancer diagnosis and therapy). To
meet the increased demand of the monoclonals, worldwide manufacturing capacity
for MAbs must increase dramatically, during the next few years, if the industry
segment is to avoid a manufacturing bottleneck. Many novel expression
strategies are being evaluated to increase the production capacity. Factors
including molecular fidelity and cost of products will be critical in the
selection of the expression systems.
Indian Journal of Biotechnology
Apoptosis: An Overview
Rana Anjum and Ashok Khar
Apoptosis, a highly ordered cascade of enzymatic
events that culminates in cell death and entails the autolytic degradation of
cellular components, is characterized by blebbing of cell membrane, nuclear and
cytoplasmic condensation, protein fragmentation and DNA degradation followed by
rapid engulfment of cell corpses by neighbouring cells. It is essential for
maintenance of cellular homeostasis and deregulation of this process leads to a
spectrum of pathological manifestations. Caspases, which form the proteolytic
network within the cell are among the critical components of cell death
process. They bring about the cleavage and degradation of a number of proteins
that result in cell demise. Mitochondria are regarded as the central control
point in the execution of apoptosis. They release a potent cocktail of
pro-apoptotic proteins into the cytosol, which results in the amplification of
the death cascade, the most prominent one being cytochrome c. The mechanism by which cytochrome c is released into the cytosol is controversial but seems to be
regulated by the Bcl-2 family of proteins.
Indian Journal of Biotechnology
Virus Resistant Transgenic Plants for
Environmentally Safe
Management of Viral Diseases
A Varma, R K Jain and A I Bhat
Plant viruses
are one of the major yield reducing factors for agricultural and horticultural
crops. In India, most destructive diseases are caused by gemini-, poty-, and
tospoviruses. Virus resistant transgenic plants (VRTPs), developed by the
transfer of transgenes from virus, plant or other origins, have been found
resistant to a wide range of viruses. The most successful approach is the viral
coat protein mediated resistance (CPMR). Other transgenes of viral origin,
which have shown promise are: replicase protein, movement protein, proteases,
and antisense sequences. ‘R’-genes from plants, plantibodies and yeast RNase
genes are also useful for developing VRTPs. In a large number of VRTPs developed
using transgenes of viral origin, resistance is conferred by
post-transcriptional gene silencing (PTGS); in some cases PTGS has been
overcome when plants are infected by a heterologous virus, indicating need for
cautious approach. Overall, the bio-safety concerns in the use of VRTPs get
insignificant, but these must be addressed scientifically. In India,
initiatives have been taken for developing VRTPs to manage important plant
viral diseases. The present world area under VRTPs is about 0.4 mha. Judging from
the success of various strategies, the area under VRTPs is expected to grow at
a fast rate in the coming years.
Indian Journal of Biotechnology
Electrostatic Interactions, Phase
Separation Behaviour and Partitioning of Proteins in Polyelectrolyte Based
Aqueous Two-Phase Systems
Vandana Gupta, Sunil Nath and Subhash Chand
Electrostatic interactions
play a major role in the purification of proteins by different methods and they
tend to be specific in nature in the presence of salts and environmental
hydrogen ion concentration (pH).
These can be readily exploited as the basis for protein isolation and purification
by aqueous two-phase system (ATPS) using a polyelectrolyte as one of the
polymeric components. Polyelectrolytes are water-soluble charged polymers and
the phase separation in the polyelectrolyte based ATPS is dependent on the
ionic composition of the system and the charge density of polymers. This review
outlines the mechanism of the phase separation in non-ionic polymers based ATPS
on the basis of water structure in polymeric solution and validity of the
hypothesis is further discussed for polyelectrolyte-based systems. Partitioning
of proteins in polyeletrolyte based ATPS is dominantly controlled by the
electrostatic interactions. Several factors that influence the partitioning of
proteins include the properties of the polymers, pH, salt type and concentration and the charge on protein
molecules. Low polymer concentration requirement for the phase separation and
high specificity of protein purification in polyelectrolyte based ATPS holds a
great promise for their large scale applications in isolation and purification
of proteins.
Indian Journal of Biotechnology
Vol 1, January 2002, pp
96-100
Development of Genetic Markers
Specific for Highly Endangered Species Rhinoceros
unicornis: Strategies and Implications in Conservation
Vipra Kapur, Anu Bashamboo and Sher Ali
Indian rhino, Rhinoceros unicornis,
is a highly endangered species and unless collective efforts are made for its
protection, the future of this species will continue to remain bleak.
Application of restriction survey and minisatellite associated sequence
amplification (MASA) for identification, cloning and sequencing of the
potential genomic fragments from this species, led to the development of
species specific DNA marker. These markers are envisaged to be useful for
ascertaining the origin of rhino's biological samples in the event of poaching
and illegal trafficking. This approach may be used for conservation and
management of other endangered species.
Indian Journal of Biotechnology
Bioinformatics: Advancing
Biotechnology through Information Technology Part I: Molecular Biology
Databases
Sudeshna Adak and Biplav Srivastava
This paper
is intended as a review of molecular biology databases and other Bioinformatics
resources available for biotechnologists aiming to use the wealth of genomic
data available today. The genomic data along with associated proteomic and
functional data are often distributed across multiple databases, requiring a
time-consuming search by the user. The explosion of information seen in
molecular biology has created a veritable maze, through which careful
navigation is required for research and innovation in biotechnology. The paper,
one of the series, introduces the readers to the major molecular biology
databases and bioinformatics tools such as BLAST for similarity searching and
RasMol for protein structure visualization. Subsequent papers will take the
readers into a journey across bioinformatics and the biotechnological
discoveries that are happening with bioinformatics. Advances in computer
technologies and the birth of the internet are also part of this revolution in
biology. Online databases have given scientists and researchers across the
world access to unimaginable volumes of biologically relevant data.
Bioinformatics, a truly multidisciplinary science, aims to use the benefits of
computer technologies in understanding the biology of life itself.
Indian Journal of Biotechnology
Biomolecular Engineering of Escherichia coli Organo-mercurial
Lyase Gene and its Expression
Imtiyaz Murtaza, Amit Dutt and Arif Ali
Studies were carried out to characterize merB gene from five wild type strains of broad-spectrum Escherichia coli, collected from five
geographically distinct regions of India. Each strain produced 23kb plasmid
from which functional merB gene
(0.64kb) was PCR amplified. The merB
gene from isolate G18, which tolerated highest concentration of organic form
(PMA) of mercury was cloned in high expression vector pQE30 and pGEMT-Easy
vector. The transformants obtained demonstrated varied results in their
appropriate hosts. The transformants (IAxpress) carrying merB gene cloned in pQE30 and negative control having pQE30 without
merB insert did not grow on agar
plates amended with 1mg/ml PMA. Due to the hyperexpression of merB in pQE30 most of the protein was found in nonfunctional
inclusion bodies and did not show any resistance as sensitive strain (Devoid of
merB gene) against PMA. On the other
hand transformants of merB cloned
pGEMT vector tolerated up to 5mg/ml of PMA, which indicates that low expression of merB in this vector produces a functional product and thus
tolerates five times more PMA than sensitive strain. The results demonstrate
that this gene can be better exploited for bioremediation of toxic form of
mercury in polluted water bodies.
Indian Journal of Biotechnology
Biotechnology—The
Interface between Agri-Resources and the Consumer
V Prakash
Indian Journal of Biotechnology, started as a quarterly journal in 2002, publishes full papers, short communications and reviews in agricultural-, animal-, environmental-, industrial-, medical-,and microbial biotechnology, bioinformatics, and socio-legal and ethical aspects in biotechnology.The latest developments in biotech-industry are covered under notes and news.