molecular biotechnology vol 2 issue 2

International Journal of

Molecular

BiotechnologyIJMBJul–Dec 2016

Mechanical Engineering

Electronics and Telecommunication Chemical Engineering

Architecture

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¬ International Journal of Thermal Energy andApplications

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« International Journal of Radio Frequency Design« International Journal of VLSI Design and Technology« International Journal of Embedded Systems and Emerging

Technologies« International Journal of Digital Electronics« International Journal of Digital Communication and Analog

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« International Journal of Solid State Materials« International Journal of Optical Sciences

Physics

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Civil Engineering« International Journal of Water Resources Engineering« International Journal of Concrete Technology« International Journal of Structural Engineering and Analysis« International Journal of Construction Engineering and

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Electrical Engineering« International Journal of Analog Integrated Circuits« International Journal of Automatic Control System« International Journal of Electrical Machines & Drives« International Journal of Electrical Communication

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Material Sciences and Engineering « International Journal of Energetic Materials« International Journal of Bionics and Bio-Materials« International Journal of Ceramics and Ceramic Technology« International Journal of Bio-Materials and Biomedical

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Chemistry « International Journal of Photochemistry« International Journal of Analytical and Applied Chemistry« International Journal of Green Chemistry« International Journal of Chemical and Molecular

Engineering

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Cell transformation

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Hidam Renubala

Ankita Singh

Deepika Bhadauria

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Commissioning Editors

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Manisha Dhoble

EDITORIAL BOARD MEMBERS

Dr. Saber Mohamed Abd-Allah Shanghai Institute of Biochemistry and Cell

Biology, CAS, China

Dr.Ashok Kumar KulkarniDepartment of Physiology, MediCiti Institute of Medical Sciences, Medchal. Secunderabad

(A. P), India

Dr. Promila SharmaDepartment of Biotechnology, Graphic Era

University, Uttarakhand, India

Dr. Brijesh PandeyAmity University, Lucknow (U.P.),

India

Dr. P Mary AnupamaDepartment of Biotechnology, ANITS,

Sangivalasa, Visakhapatnam (A.P.), India

Dr. Biswajit Majumdar Calcutta University Institute of Post Graduate Medical Education and Research [IPGMER]

/University College of Medicine [UCM], Kolkata, India

Dr. Indraneel GhoshManager Systems Biology Department Sun Pharmaceuticals Advanced Research Centre

Limited, Vadodara (Gujarat), India

Dr. Arvind Diwan(Marine Fisheries), ICAR Project Director

(Biotechnology), Mahatma Gandhi Mission Institute of Health Sciences (Deemed

University), Aurangabad (Maharashtra), India

Dr. Ir. Sreenivas Reddy BathulaDepartment of Biotechnology, Bapuji Institute of

Engineering & Technology, Karnataka, India

Dr. Rajat BanerjeeDepartment of Biotechnology, University of

Calcutta, India

Dr. Devarakonda Srinivasa Rao Department of Biotechnology Acharya

Nagarjuna University, Guntur (DT), India

Dr. N. Manikanda BoopathiDepartment of Fruit Crops, Horticultural

College and Research Institute, Tamil Nadu Agricultural University, Tamil Nadu, India

Dr. Avinash CheekothManagement of the Scientific Centers,

Department of the President's Affairs, UAE

Dr. Aumreetam Dinabandhu ICAR - NRC on Plant Biotechnology,

New Delhi, India

Dr. Sanjay Mahendrakumar Dave Department of Biotechnology,

Hemchandracharya North Gujarat University, Gujarat, India

Dr Indraneel GhoshSystems Biology Department,

Sun Pharmaceuticals Advanced ResearchCentre Limited, Vadodara, Gujarat, India

Dr. Sukhjeet KaurDepartment of Biotechnology, SUS College of Engineering & Technology, Tangori-Mohali,

India

Dr. Sukhadev Bhaskar NandeshwarCentral Insititute for Cotton Research,

Nagpur, India

EDITORIAL BOARD MEMBERS

Dr Vara PrasadNano Research for Advanced Materials,

Bangalore, India

Dr. Anjali PriyadarshiniDepartment of Biotechnology, PGIMER,

Chandigarh, India

Dr. Soma RoyDepartment of Biotechnology, CBIT, Hyderabad,

India

Dr. Tha. ThayumanavanSchool of Biotechnology, Dr.G.R. Damodaran

College of Science, Coimbatore, India.

Dr. K.S. KarthikeyanEndocrinology and Metabolism Division,

National Institute of Nutrition, Indian Council of Medical Research, Hyderabad, India.

Dr. Aseel Mahmood AljamaliSurgery Department, College of Medicine,

Kufa University, Iraq

Dr. M. Anusuyadevi JayachandranMolecular Gerontology Laboratory, Department

of Biochemistry, Bharathidasan University, Tamil Nadu, India

Dr. Rekha KhandiaDepartment of Science and Technology,

High Security Animal Disease Laboratory,IVRI, ICAR, Bhopal (MP), India

Dr. Kumaresan MohanUdaya School of Engineering

Tamil Nadu, India

Dr. Subha ParameshwaranDepartment of Microbiology, KJ Somaiya

Medical College, Sion, Mumbai, India

Dr. Guru PrasadBasavatarakam Indo-American Cancer Hospital

and Research Institute, Hyderabad, India

Dr. Shashwat SharadDepartment of Biotechnology, Dr YS Parmar University of Horticulture & Forestry, Solan

(HP), India

Dr. S. Thangminlal VaipheiRadiation and Molecular Biology Unit,

Department of Biochemistry, North-Eastern Hill University (NEHU), Meghalaya, India

Dr. Kakali PurkayasthaScientist, AIIMS, New Delhi, India

Dr. J. John Peter PaulSt. Xavier's College (Autonomous),

Tamil Nadu, India

From the Editor's Desk

Dear Readers,

We would like to present, with great pleasure, the inaugural volume of a new scholarly

journal, International Journal of Molecular Biotechnology. This journal is part of the

Applied Sciences, and is devoted to the scope of present Molecular Biotechnology issues,

from theoretical aspects to application-dependent studies and the validation of emerging technologies.

This new journal was planned and established to represent the growing needs of Molecular Biotechnology

as an emerging and increasingly vital field, now widely recognized as an integral part of scientific and

technical investigations. Its mission is to become a voice of Molecular Biotechnology, addressing

researchers and practitioners in this area.

The core vision of International Journal of Molecular Biotechnology in JournalsPub is to propagate novel

awareness and know-how for the profit of mankind ranging from the academic and professional research

societies to industry practitioners in a range of topics in Molecular Biotechnology in general. JournalsPub

acts as a pathfinder for the scientific community to publish their papers at excellently, well-timed &

successfully. International Journals of Molecular Biotechnology focuses on original high-quality research

in the realm of Cell transformation, Gene cloning systems, Next generation sequencing, Molecular and cell

biology automation, Protein purification and analysis, Transgenic species, Therapeutic proteins, Cells

genetic and metabolic capacity, Molecular markers etc.

The Journal is intended as a forum for practitioners and researchers to share the techniques of Molecular

Biotechnology and solutions in the area. Many scientists and researchers have contributed to the creation

and the success of Molecular Biotechnology. We are very thankful to everybody within that community who

supported the idea of creating an innovative platform. We are certain that this issue will be followed by many

others, reporting new developments in the field of Molecular Biotechnology.

This issue would not have been possible without the great support of the Editorial Board members, and we

would like to express our sincere thanks to all of them. We would also like to express our gratitude to the

editorial staff of JournalsPub, who supported us at every stage of the project.

It is our hope that this fine collection of articles will be a valuable resource for Molecular Biotechnology

readers and will stimulate further research into the vibrant area of Molecular Biotechnology.

Puneet Mehrotra

Managing Director

1. Diagnosis of Forest Pathogens: A Molecular Approach – A Review S. Rawat, A. Bagwari, Y.P. Singh 1

2. Breeding for Disease Resistance in Crops by Molecular Method Archana Bagwari 8

3. Molecular Markers in Livestock ImprovementSanjay Kumar 14

4. Production of Therapeutic Proteins: A ReviewAmandeep Kushan 20

5. Rapid DNA Isolation Protocol for ISSR and SSR Markers Analysis in Trans-Himalayan Cabbage SpeciesS. Allen, S. Kapoor, M.K. Patel, V. Kumar, O.P. Chaurasia, B. Kumar 26

Contents

IJMB (2016) 1–7 © JournalsPub 2016. All Rights Reserved Page 1

International Journal of Molecular Biotechnology Vol. 2: Issue 2

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Diagnosis of Forest Pathogens: A Molecular Approach – A Review

S. Rawat*, A. Bagwari, Y.P. Singh

Forest Pathology Division, FRI, Dehradun, India

ABSTRACT

In India forest is the main source of timber and livelihood for many farmers. But the condition of forestry is not good in India as many factors affect their production. One of the main factor is pathogens especially fungi. Many wood rotting fungi are reported to cause mortality in important forestry trees in India. Accurate identification of pathogen is an important strategy to initiate curative measures. Diagnosis of any disease is the first stage of its management. In recent time, researcher use many new methods, as manual methods for the detection of pathogens is time consuming and needs expert supervision. These limitations led to the development of molecular approaches due to high accuracy and specificity. Molecular technique will be effective for the detection of pathogens. By using molecular approaches forest pathogens can be managed easily. So, aim of this review is to study the role of molecular methods in the management of forest pathogens. Keywords: diagnosis, fungi, molecular markers INTRODUCTION Forest is affected by various biotic agents or there are several agents which cause disease on tree. For example, bacteria, virus, protozoans, fungi, etc. Fungi cause the majority of forest diseases (Edmonads, et al., 2010). Some examples of fungal disease which causes death of the plants are, wilt disease of Dalbergia sissoo Roxb. Caused by

Fusarium solani (Bagchee, 1945; Bakshi, 1954), Ganoderma root rot caused by Ganoderma lucidum on various tree, damping off by Rhizoctonia solani, leaf spot and leaf blight by various fungal pathogens. Further, wood decaying fungi are the important elements of forest ecosystems, as they take Care of the decomposition of coarse woody debris (fallen trunks, branches, stumps, etc.).



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Breeding for Disease Resistance in Crops by Molecular Method

Archana Bagwari*

Forest Pathology Division, FRI, Dehradun, India

ABSTRACT

Tree/crops are the prime source of food in India. Crops also contribute to the national exchequer. At the same time, plants are the reservoir of many organisms (microorganism, nematodes, insect, etc.) as they have rich source of nutrient in it. These organism cause disease on plants and sometimes caused death of the plants. So, it can create severe food threat in our system. Plants lack immune system to fight against such disease but have a defense mechanism. Plant pathologists discover various methods for the control of the disease. Breeding for disease resistance is one of the best examples to overcome against severe pathogen. It is very quick methods for disease resistance. Keywords: disease, pathogens, resistance INTRODUCTION Plant disease is a serious problem for plant pathologist. Although, many advance technique have been developed to control the disease, but our universal food supply is still vulnerable by a several pathogens and pests (Moffat, 2001; Garelik, 2002 and Many authors, 1999). Diseases caused by the pathogens can greatly reduce crop outturn and the effect of disease outburst is mainly acute in developing nations like in India. Oerke (2006) reported that an average of 26% of the worldwide crop yielding is lost each year due to pre-harvest pests and pathogens. Chemicals (Fungicides, pesticides, etc.) provide effective protection but their suitableness can be affected by adverse environmental conditions and by the emergence of resistant pathogen strains (McDowell and Woffenden, 2003) [1–5]. Expanding rate of human population, loss of farming area because of environmental change, disintegration, for example, those

created by pathogens however much as could be expected (Bebber et al., 2013). The four major crop yielded universally, are rice (which feeds more than half the world population), wheat, maize and banana are continuously exposed by destroying emerging infectious diseases in both developing and developed countries (Fisher et al., 2012). Magnaporthe oryzae caused Rice blast is one of the most destructive diseases of the rice in several part of the world (Skamnioti and Gurr, 2009; Helliwell and Yang, 2013; Helliwell et al., 2013). Another serious threat to food security is Basidiomycetes fungus Puccinia graminis f. sp. tritici (the causal agent of wheat stem rust). Another major fungi, the oomycete pathogen Phytophthora infestans is the causative agent of late blight of potato, resulting in the deaths of 1.25million people during the Irish potato famine of 1845 (Haverkort et al., 2008; Pennisi,2010) (Figure 1, Table 1) [6–10].



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Molecular Markers in Livestock Improvement

Sanjay Kumar Central University of Rajasthan, Ajmer, India

BACKGROUND Recent advancements in DNA technologies have made it conceivable to reveal an extensive number of genetic polymorphisms at the DNA sequence level, and to utilize them as markers for assessment of the genetic basis for the watched phenotypic variability. The markers revealing varieties at DNA level are alluded to as the molecular markers. Based on techniques utilized for location, these markers are arranged into two noteworthy classifications: Hybridization-based markers PCR-based markers The molecular markers have interesting genetic properties and methodological advantages that make them more helpful and manageable for genetic examination contrasted with other genetic markers. The conceivable uses of molecular markers in domesticated animals change have been surveyed with reference to routine and transgenic reproducing procedures. In breeding strategies, molecular markers have a several short-range or immediate applications (viz. parentage determination, genetic distance estimation, determination of twin zygosity and free martinism, and sexing of pre-implantation embryos and identification of disease carrier) and long-range applications (viz. gene mapping, and marker-assisted selection) and long-go applications (viz. quality mapping, and marker-helped choice) [1, 2] In transgenic breeding, molecular markers can be utilized as reference focuses for identification, isolation and manipulation

of the relevant genes, and for identification of the animals carrying the transgenes. The advance being developed of molecular markers proposes their potential use for genetic change in domesticated animal’s species. The advance in recombinant DNA technology and gene cloning during the most recent two decades has gotten revolutionary changes in the field of fundamental and additionally of connected genetic by giving a few new ways to deal with genome analysis with more prominent genetic resolution. It is currently conceivable to reveal countless polymorphisms at the DNA sequence level, and to utilize them as markers for assessment of the genetic reason for the watched phenotypic variability [3]. Despite the fact that hypothetically DNA sequencing is the immediate way to deal with uncovers such DNA polymorphism, it has two reasonable impediments:

Sequencing needs initial cloning of (i)the quality or DNA fragment at which allelic variety of intrigue exists, and

(ii) It requires appropriate and savvy technique for scoring DNA sequence variety. In any case, the aberrant approach for revealing of genetic variation at the DNA level utilizing molecular or DNA markers blocks the above restrictions. Since the primary show of DNA-level polymorphism, known as the limitation fragment length polymorphism (RFLP), a practically boundless number of molecular markers have aggregated. Currently, more



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Production of Therapeutic Proteins: A Review

Amandeep Kushan Department of Biotechnology, Mewar University, Chittorgarh, Rajasthan, India

BACKGROUND Therapeutic proteins will be proteins that are engineered in the lab for pharmaceutical utilize. Insulin was the main remedial protein to be acquainted with treat diabetes in the 1920s. Because of recombinant DNA-technology proteins can be produced in particular host cells (e. g. bacteria, yeast or mammalian cells) under characterized conditions. Contingent upon the necessities of the protein the production host must be picked precisely. Mammalian cells are the primary host of decision in light of the fact that their posttranslational changes, for example, glycosylation and sialylation has the greatest effect on the protein's pharmacokinetics and effectiveness. Protein-based treatments are utilized for instance as a part of the treatment of: Cancer Infectious diseases Hemophilia Anemia Multiple sclerosis Hepatitis B/C Therapeutic proteins allow an individualized treatment approach by supporting a particularly focused on Therapeutic process by remunerating the lack of an essential protein. The quickest developing class of Therapeutic proteins is antibodies. From one perspective they bolster the safe reaction and then again they can prompt the aggregation of Therapeutic agents at a

tumor site through conjugation/fusion [1, 2]. Antibodies are Therapeutic proteins used for the treatment of malignancy as well as auto-immune diseases. Production of Therapeutic Proteins by Gene Transfer There are numerous proteins fundamental to great wellbeing that a few people can't produce due to genetic defects. These proteins incorporate different blood-clotting components bringing about hemophilia, insulin (bringing about diabetes), development hormone (bringing about absence of appropriate development), and different proteins, the organization of which revises neurotic conditions or results in other Therapeutic advantages. The early work in this field utilized bacteria. Some bacteria in a bacterial culture may contain little round DNA particles called plasmids. These plasmids are not part of the chromosomal DNA that is controlled that is possessed by all the bacteria of the culture. As these outstanding bacterial cells replicate by binary fission or cell division, the plasmids are transmitted to the Daughter cells. They can likewise be transmitted to different cells by conjugation. Researchers have figured out how to use plasmids to exchange human genes to bacterial cells. If the gene inserted into the plasmid of bacteria into the human gene for insulin, for instance, the bacteria into which this gene is inserted produces human insulin.

IJMB (2016) 26-31© JournalsPub 2016. All Rights Reserved Page 26


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Rapid DNA Isolation Protocol for ISSR and SSR Markers Analysis in Trans-Himalayan Cabbage Species

S. Allen, S. Kapoor, M.K. Patel, V. Kumar, O.P. Chaurasia*, B. Kumar

Department of Plant Biotechnology, Defense Institute of High Altitude Research DRDO, Chandigarh, India

ABSTRACT

In the present study, we present a simple one day protocol for DNA extraction from cabbage, with no requirement of RNase and overnight precipitation with isopropanol. Due to high endogenous phenolics and polysaccharides and other substances in plants, it is difficult to isolate DNA with high concentration and optimum ratio. With added protein and RNA contamination, ISSR and SSR based primer amplification pose a challenge for the same. This method involves modified CTAB protocol with fine crushing by liquid nitrogen. The reagents required during the extraction are CTAB, NaCl, EDTA, Tris HCl and Ascorbic acid. The extraction buffer should be freshly prepared on the same day as results indicated low yield with overnight extraction buffer. The yield of DNA isolated varies from 107.2 to 3855 ng/µl with ratio ranging from 1.79 to 1.88. The primers developed for the case study were mainly ISSR (inter Simple Sequence Repeats) and SSR (Simple sequence Repeats) which were first standardized for PCR reaction mixture such as MgCl2 (1.8 mM), primers (0.2µM) and Taq Polymerase (0.3 units) with 20 ng/µl of DNA. The gradient PCR showed the annealing temperature of all the primers ranging from 55 to 64°C. Polymorphism was seen among all the amplified PCR products run on 2% agarose gel electrophoresis. Keywords: cabbage, CTAB, DNA extraction, ISSR, polymorphism Abbreviations: CTAB, cetyltrimethyl ammonium bromide; EDTA, ethylene diaminetetraacetic acid; NaCl, sodium chloride; TrisHCl, tris hydrochloride INTRODUCTION Plants produce secondary metabolites that interfere not only with extraction of high quality genomic DNA but also with the subsequent reactions such as PCR [1, 2]. PCR based DNA marker analysis is now-a-days, widely used in several crop plants for genetic diversity analysis, Marker assisted selection, (MAS), genetic mapping and varietal identification [3]. An array of DNA isolation protocols are available and are being used for the extraction of nucleic acids from plants [4]. RNA and protein contamination in DNA may result in no band or smear formation during PCR amplification. DNA

amplification can be affected by the presence of certain impurities/ co purifying inhibitors, which reduces PCR efficiency. Therefore quality of DNA is of utmost importance [5–7]. The family Brassicaceae consists of approximately 340 genera and more than 3350 species [8]. In addition to providing vegetables, fodder, vegetable oil and condiment, Brassicas are important sources of dietary fiber, vitamins and anticancer compounds [9]. Cabbage (Brassica oleracea L. varcapitata), family


Molecular

BiotechnologyIJMBJul–Dec 2016

Mechanical Engineering

Electronics and Telecommunication Chemical Engineering

Architecture

Office No-4, 1 Floor, CSC, Pocket-E,Mayur Vihar, Phase-2, New Delhi-110091, India

E-mail: [email protected]

¬ International Journal of Thermal Energy andApplications

¬ International Journal of Production Engineering¬ International Journal of Industrial Engineering

and Design¬ International Journal of Manufacturing and

Materials Processing¬ International Journal of Mechanical Handling and

Automation

« International Journal of Radio Frequency Design« International Journal of VLSI Design and Technology« International Journal of Embedded Systems and Emerging

Technologies« International Journal of Digital Electronics« International Journal of Digital Communication and Analog

Signals

« International Journal of Housing and Human SettlementPlanning

« International Journal of Architecture and InfrastructurePlanning

« International Journal of Rural and Regional PlanningDevelopment

« International Journal of Town Planning and Management

Applied Mechanics

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Computer Science and Engineering « International Journal of Wireless Network Security« International Journal of Algorithms Design and Analysis« International Journal of Mobile Computing Devices« International Journal of Software Computing and Testing« International Journal of Data Structures and Algorithms

Nanotechnology« International Journal of Applied Nanotechnology« International Journal of Nanomaterials and Nanostructures« International Journals of Nanobiotechnology

« International Journal of Solid State Materials« International Journal of Optical Sciences

Physics

« International Journal of Renewable Energy and itsCommercialization

« International Journal of Environmental Chemistry« International Journal of Agrochemistry« International Journal of Prevention and Control of Industrial

Pollution

Civil Engineering« International Journal of Water Resources Engineering« International Journal of Concrete Technology« International Journal of Structural Engineering and Analysis« International Journal of Construction Engineering and

Planning

Electrical Engineering« International Journal of Analog Integrated Circuits« International Journal of Automatic Control System« International Journal of Electrical Machines & Drives« International Journal of Electrical Communication

Engineering« International Journal of Integrated Electronics Systems and

Circuits

Material Sciences and Engineering « International Journal of Energetic Materials« International Journal of Bionics and Bio-Materials« International Journal of Ceramics and Ceramic Technology« International Journal of Bio-Materials and Biomedical

Engineering

Chemistry « International Journal of Photochemistry« International Journal of Analytical and Applied Chemistry« International Journal of Green Chemistry« International Journal of Chemical and Molecular

Engineering

« International Journal of Electro Mechanics andMechanical Behaviour

« International Journal of Machine Design andManufacturing

« International Journal of Mechanical Dynamicsand Analysis

« International Journal of Fracture and damageMechanics

« International Journal of Structural Mechanicsand Finite Elements

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Biotechnology « International Journal of Industrial Biotechnology and

Biomaterials« International Journal of Plant Biotechnology« International Journal of Molecular Biotechnology« International Journal of Biochemistry and Biomolecules« International Journal of Animal Biotechnology and

Applications3 more...

Nursing « International Journal of Immunological Nursing« International Journal of Cardiovascular Nursing« International Journal of Neurological Nursing« International Journal of Orthopedic Nursing« International Journal of Oncological Nursing

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