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

        Navin Khanna

 

 

 

Apoptosis: an overview

58

        Rana Anjum & Ashok Khar

 

 

Virus resistant transgenic plants for environmentally safe management of viral diseases

73

        A Varma, R K Jain & A I Bhat

 

 

Electrostatic interactions, phase separation behaviour and partitioning of proteins in polyelectrolyte based aqueous two-phase systems

87

        Vandana Gupta, Sunil Nath & Subhash Chand

 

Development of genetic markers specific for highly endangered species, Rhinoceros unicornis: strategies and implications in conservation

96

        Vipra Kapur, Anu Bashamboo & Sher Ali

 

 

Bioinformatics: advancing biotechnology through information technology Part I: molecular biology databases

101

        Sudeshna Adak & Biplav Srivastava

 

 

Biomolecular engineering of Escherichia coli organomercurial lyase gene and its expression

117

        Imtiyaz Murtaza, Amit Dutt & Arif Ali

 

 

Biotechnology—the interface between agri resources and the consumer

121

        V Prakash

 

 

 

Instructions to Contributors

126

 

 

Indian Journal of Biotechnology

Vol 1, January 2002, pp 9-16

 

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

Vol 1, January 2002, pp 17-28

 

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

Vol 1, January 2002, pp 29-38

 

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

Vol 1, January 2002, pp 39-49

 

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

Vol 1, January 2002, pp 50-57

 

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

Vol 1, January 2002, pp 58-72

 

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

Vol 1, January 2002, pp 73-86

 

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

Vol 1, January 2002, pp 87-95

 

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

Vol 1, January 2002, pp 101-116

 

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

Vol 1, January 2002, pp 117-120

 

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

Vol 1, January 2002, pp 121-125

 

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.