eChapters

Introduction

Western blot analysis can detect one protein in a mixture of any number of proteins while giving you information about the size of the protein. It does not matter whether the protein has been synthesized in vivo or in vitro. This method is, however, dependent on the use of a high-quality antibody directed against a desired protein. So you must be able to produce at least a small portion of the protein from a cloned DNA fragment. You will use this antibody as a probe to detect the protein of interest.

Western blotting tells you how much protein has accumulated in cells. If you are interested in the rate of synthesis of a protein, Radio-Immune Precipitation (RIP) may be the best assay for you. Also, if a protein is degraded quickly, Western blotting won’t detect it well; you’ll need to use (RIP). See the section on RIP for more information, as well as a helpful comparative chart that illustrates the differences between these two techniques.

Abstract

Bioinformatics is an important interdisciplinary field that brings together several areas of science, including molecular biology, biostatistics, and computer science (vis-a-vis information technology). With the rapid increase in the sequencing of DNA/RNA/protein molecules, as well as the growing understanding of the interconnections between different areas of biological science, the importance of bioinformatics is only increasing. This chapter seems to provide a thorough introduction to the definition, concept, and importance of bioinformatics. It also appears to highlight the relevance and potential scope of the subject for students interested in pursuing a career in the field. As bioinformatics continues to evolve and expand, it is likely to become an increasingly important area of research and development in the life sciences.

What is Nanotechnology? Answers differ depending on who you ask, and their background. Broadly speaking however, nanotechnology is the act of purposefully manipulating matter at the atomic scale, otherwise known as the “nanoscale.” Coined as “nano-technology” in a 1974 paper by Norio Taniguchi at the University of Tokyo, and encompassing a multitude of rapidly emerging technologies, based upon the scaling down of existing technologies to the next level of precision and miniaturization. Taniguchi approached nanotechnology from the ‘top-down’ standpoint, from the viewpoint of a precision engineer.

Foresight Nanotech Institute Founder K. Eric Drexler introduced the term “nanotechnology” to the world in 1986, using it to describe a ‘bottom-up’ approach. Drexler approaches nanotechnology from the point-of-view of a physicist, and defines the term as “large-scale mechano synthesis based on positional control of chemically reactive molecules.” In the future,“ nanotechnology” will likely include building machines and mechanisms with nanoscale dimensions, referred to these days as Molecular Nanotechnology (MNT).

Wheat also known as bread wheat (Triticum aestivum L. em. Thell, 2n = 6x = 42, family Poaceae formerly Gramineae or grass family, a self-pollinated crop), second only to rice as source of calorie has been grown on 218 million ha with production of 765 million tons globally during 2019-20. Top five countries, namely, China (135 million tons), India (103 million tons), Russia (77 million tons), USA (51 million tons) and Canada (34 million tons) account for 65% of the global production. Other major countries growing wheat are Ukraine (28 million tons), Pakistan (26 million tons), Australia (24 million tons), Argentina (21 million tons) and Turkey (18 million tons). Wheat is eaten by 2.5 billion people in 89 countries and distributed from Scandinavia to South America and across Asia. Global annual wheat trade is worth USD 50 billion.

Agriculture Scenario

It has been seen since last few decades there is huge decrease in the production of the agricultural production. Three factors which can be held responsible for the increase of agricultural production per hectare:

1. Overall increase in efficiency due to the merger of farms and a more efficient redistribution of farm land.
2. More efficient cultural practises and better quality and availability of inputs (fertilizer, pesticides).
3. Genetic improvement of crops.

At this moment over 6 billion people need to be fed and it is expected that in the year 2050 more than 10 billion people will inhabit the earth (FAO, 1996). Moreover, demands per capita will rise when standards of living in developing counties improve. As a consequence there will be a huge increase in the demand for food, and production will need to triple in the coming 40 years. A continued effort to secure agricultural production in the future will therefore be vital. Reduction of losses caused by a-biotic and biotic stress will be a key issue. Scenario studies on world food security generally assume that a large increase in production will be achieved by genetic improvement of crop species and biotechnology. History has indeed shown a continuous increase in crop yields, resulting from plant breeding efforts. However, in the light of the speed at which the human population develops, and taking into account the expected reduction of available arable land due to climate change and human intervention, it may be necessary to accelerate the rate at which genetic improvement is achieved. Modern biotechnology provides new tools that can facilitate development of improved plant breeding methods and augment our knowledge of plant genetics. The knowledge that is obtained with these new tools can be used to contribute to an enhanced food security throughout the world.

World Intellectual Property Organization

http://www.wipo.int,http://www.wipo.org

The World Intellectual Property Organization - referred to in abbreviated form as WIPO in English, and OMPI in French and Spanish - was established by a convention signed at Stockholm on July 14, 1967, and entitled “Convention Establishing the World Intellectual Property Organization.” The WIPO Convention entered into force in 1970. WIPO became a specialized agency in the United Nations system of organizations in 1974 and dedicated to the use of intellectual property (patents, copyright, trademarks, designs, etc.) as a means of stimulating innovation and creativity.

WIPO promote the development and use of the international IP system through Services, Law, Infrastructure and Development.

12.1 Introduction

“Precision farming,” also known as site-specific management, describes a bundle of new information technologies applied to the management of large-scale, commercial agriculture. Precision farming technologies include, for example: personal computers, satellite-positioning systems, geographic information systems, automated machine guidance, remote sensing devices and telecommunications. If they function as designed, ubiquitous wireless sensors will become an essential tool for bringing this vision of precision farming to maturity.

Abstract

The two most important tasks for a patentee to undertake after being granted a patent, in any patent regime, would be to pay the renewal fees necessary to keep the patent alive, and provide proof of having worked the patent in the jurisdiction. This article seeks to highlight features of India’s legal requirements with regard to the working of patents on a commercial scale. The number of patents granted in a country is an indicator of the technological strength of the country. However, it should be noted that actual technological growth depends upon the working of patents. Working of a patent means the commercial exploitation of the invention that is embodied in the patent.

Objectives

• Chi-square test for goodness of fit of genetic ratios.

Introduction

Karl Pearson developed the chi square test of statistical significance which is commonly used in Mendelian and population genetics. Chi square (χ²) test is a test of statistical significance which is used to test the significance difference between observed and expected frequencies or ratios. The purpose of χ² test is to decide if a set of observed data is according to an expected ratio or if it agrees well with an expected or theoretical distribution.

The chi square test of independence allow us to evaluate whether the segregation of alleles at one locus is independent of the segregation of alleles at another locus. To distinguish between inheritance patterns that obey Mendel’s laws versus those that do not, a conventional strategy is to make crosses and then quantitatively analyze the offspring’s. Based on the observed outcome, an experimenter may make a tentative hypothesis. For example, it may seem that the data are obeying Mendel’s laws. Hypothesis testing provides an objective, statistical method to evaluate whether the observed data really agree with the hypothesis.

The underlying principles behind a statistical approach is to evaluate the goodness of fit between the observed data and the data that are predicted from a hypothesis. This is sometimes called a null hypothesis because it assumes there is no real difference between the observed and expected values. A high deviation would be found between the observed and expected values. If this occurs, the hypothesis is rejected. Hopefully, the experimenter can subsequently propose an alternative hypothesis that has a better fit with the data.

The null hypothesis is a particular claim concerning how the data is distributed. More will be said about the construction of the null hypothesis later. The null and alternative hypotheses for each chi square test can be stated as

Objectives

• Study of gene mapping.

Introduction

All the genes that are linked together form a linkage group. Genes of a linkage group may be presented on a single straight line in the same order in which they are normally present on the concerned chromosome. The number of different linkage groups in a species is, as a rule, equals to its gametic chromosome number (n). The distance between two neighbouring genes is proportional to the frequency of recombination (%) between them depicted on a line drawing. Such a line drawing depicting the linked genes the recombination frequencies between them is known as Linkage map, Genetic map or Chromosome map. Linkage maps provide information on the genes that are linked together and the frequencies of recombination that may be expected between them. The frequency of recombination between two linked genes cannot exceed 50% which is the frequency in case of independent segregation.

Objectives

• Better understand the law of independent assortment.

History

The law was proposed by Gregor Johann Mendel with the other laws together called as ‘Mendelian laws of inheritance’, in 1866. He used pea plant as a model organism, and studied 7 characters.

Principle

The Law of Independent Assortment, also known as “Inheritance Law”, states that alleles of different genes assort independently of one another during gamete formation. Independent assortment for two genes can be explained by assuming that the two genes are located in two different chromosomes. The two alleles of a gene will be located in the two homologues of the concerned chromosome. Independent separation of these two pairs of chromosomes at anaphase I of meiosis will lead to the independent segregation of the genes located in them. Thus any allele of one gene is equally likely to combine with any allele of the other gene and pass into the same gamete. Independent segregation of two genes produces four different types of gametes in equal proportion. A random union among these gametes gives rise to sixteen possible zygotes. These zygotes yield a 9:3:3:1 phenotypic ratio, which is known as the typical dihybrid ratio. When two pairs of independent alleles enter into F₁ combination, both of them have their independent dominant effect. These alleles segregate when gametes are formed but the assortment occurs independently at random and quite freely.

Objectives

• Detection of linkage using Chi-square test for goodness of fit.

Introduction

The tendency of two or more genes to stay together during inheritance is known as linkage and it is the consequence of the concerned genes being located in the same chromosome. Linked genes do not show independent segregation, as a result the ratios obtained in F₂ and test cross generation are significantly different from the expected ratios of 9:3:3:1 and 1:1:1:1, respectively.

Detection of linkage χ² using test

In genetic experiments, when the material under study is segregating simultaneously for two characters, it may be of interest to know whether the two characters are inherited independently or they are linked. The χ² test can be used for this purpose. χ² test is used in detecting linkage in back crosses and inter crosses.

First the χ² is computed based on the theoretical segregating ratios. If χ² is not significant it may be concluded that the loci segregate independently. If χ² is significant we may suspect linkage between the loci. The χ² is then partitioned into components assignable to different factors.

Objectives

• Better understand the phenomenon of multiple alleles.

Introduction

The word allele is a general term to denote the alternative forms of a gene or contrasting gene pair that denote the alternative form of a gene is called allele. These alleles were previously considered by Bateson as hypothetical partner in Mendelian segregation. Mendel and his followers used the term ‘allele’ or ‘alleleomorph’ to denote the alternative form of the normal gene. It means the genes for tall and dwarf characters of pea plant are alleles. The former is normal or wild allele and the later is mutant allele. A gene can mutate several times producing several alternative expressions. When three or more alleles are found for any particular gene, these are called ‘multiple alleles’. These occupy the same locus in homologous chromosomes.

Multiple alleles are defined as genes that are members of the same gene pair and are located on the same locus. All of them control the same character but each of the allele affects that character somewhat differently than the others. In any diploid organism, a particular gene pair is represented by two alleles of the series, one allele on each of the two chromosomes of a homologous pair. This is because an organism has just two chromosomes of each type. These form a homologous pair. However, in the individuals of a population or species, many alternative forms of the same gene may be present. The grouping of all the possible alleles of a gene pair is defined as a system of multiple alleles and their mode of inheritance is called multiple allelism. The multiple allele system can be represented as A1, A2, A3, A4 etc.

Objectives

• Better understand the phenomenon of probability.

Introduction

The concept of probability is basis to many principles of classical genetics (segregation, independent assortment etc.) The principals of population genetics are based on the considerations of laws of probability. Probability is the likelihood of occurrence of an event.

Trial

A trial refers to an opportunity provided for an event to occur, e.g. tossing of a coin once constitute one trial as it provides an opportunity for head or tail to occur. A new borne can be either a boy or a girl.

Storage

Nucleic acids are very sensitive to contaminations hence care should be taken while processing and storing DNA and RNA. The following properties of reagents and conditions need to be taken care of:

Heavy metals cause breakage of phosphodiester linkage. One can use EDTA which is an excellent heavy metal chelator.

UV rays cause extensive damage to nucleic acids. UV light at 260 nm causes breakage of thymine dimers and cross-link, including loss of biological activities.

Ethidium bromide results in photo oxidation of DNA. Oxidation products can cause phosphodiester breakage.

Nucleases are found on human skin; therefore, it is advisable to avoid direct or indirect contact between nucleic acids and fingers.

Most DNAes are not very stable; however, many RNAses are very stable and can absorb to glass or plastic and remain active.

At -20°C: extensive single and double strand breaks results. Hence for long-term storage it should be stores at -70°C. ? It is best to store DNA in high salt ( >1M) in the presence of high EDTA (>10mM) at pH 8.5, for long-term storage.

Storage of DNA in CsCl with ethidium bromide in the dark is an excellent storage medium. There is about one phosphodiester break per 200 kb of DNA per year.

Storage of ë DNA in the phage is better than storing the pure DNA.

Cell Genotypes

A large variety of bacterial strains, like E. coli, are frequently used in molecular biological experiments. Students should choose the strain or strains required for an experiment on the basis of genetic properties of the strains. It is thus pertinent that a sound knowledge of the genetic makeup of the E. coli stains is acquired.

The genetic makeup of the strains of E.coli is distinctly different from the wild type (K12 or B strains) strains from which they are derived and contains genes that can be modified. Genes are supposed to be mutated when they are not listed in a genotype. In most cases, the genes listed are mutated to the point of inactivity. In a few cases, a “-” (indicating inactive or absent or deleted), “+” (indicating active). , The genotype is a factor in choosing a bacterial strain. Chosen strains should be ascertained for their utility for the specific experiment.

Small Scale Cultures

Fresh culture from a freshly streaked plate should be taken while experimenting with E. coli cells. For preparing a small scale culture of E. coli , prepare 3-5 ml of LB medium/broth (or appropriate broth include antibiotic if the culture contains a plasmid) in two sterile 50 ml tubes. Inoculate one tube with a single colony from a fresh plate. The second tube is used as a broth control. Incubate both tubes at 37°C, shaking vigorously overnight. Inspect the tubes the next morning. The broth control should be clear and the inoculated culture should be very turbid. Make a note of any debris found in the tubes and only incubate longer if the culture is not dense. Never allow cells to overgrow. For some applications, cells can be stored at 4°C for short periods prior to use.

ABSTRACT

This paper provides an overall view on the history, structure and functions of World Intellectual Property Rights Organization.

Introduction

Human genius is the source of all works of art and invention. These works are the guarantees of a life worthy of men.  It is duty of the state to ensure with diligence the protection of the arts and inventions,” reads an inscription on the copula in the WIPO Headquarters building in Geneva. True to this inscription, WIPO functions through various mechanisms fostering judicious utilization of human intellect.  This paper intends presenting an overview on WIPO.

X-ray scattering techniquesarea family of non-destructive analytical techniques which reveal information about the crystallographic structure, chemical composition, and physical properties of materials and thin films. These techniques are based on observing the scattered intensity of an X-ray beam hitting a sample as a function of incident and scattered angle, polarization, and wavelength or energy.

Wilhelm Conrad Roentgen, German physicist, born on November 8, 1895 discovered X-rays and was awarded the first ever Nobel Prize in Physics in 1901 in recognition of the extraordinary services he has rendered by the discovery of the remarkable rays subsequently named after him. These short-wave electromagnetic radiations have the wavelengths that are matching with both the atomic sizes and shortest interatomic distances. But, the index of refraction of X-rays is near unity for all materials and hence they could not be focused by a lens in order to observe small objects such as atoms, as it is done by glass lenses in a visible light microscope or by magnetic lenses in an electron microscope. Thus, in general, X-rays cannot be used to image individual atoms directly. As an alternative to this difficulty, Max von Laue in 1912 proved that the periodicity of the crystal lattice allows atoms in a crystal to be observed with exceptionally high resolution and precision by means of X-ray diffraction for the first time by using a single crystal of hydrated copper sulfate (CuSO₄ 5H₂O).

Zerovalent (nZVI) Ionic Compounds

Although Ag⁺ contains a filled d¹⁰ shell, the structures of Ag compounds can in many instances be understood only if one assumes the existence of weak attractive Ag⁺ - Ag⁺ interactions. Similar attractive forces have been shown to be responsible for the formation of gold cluster compounds. In Ag⁺ solid state compounds the unoccupied 5s and 5p orbitals give rise to empty bands which may be partially occupied thus resulting in subvalent Ag compounds; examples are Ag₃O and Ag₂F. The compound Ag₅SiO₄ prepared from Ag and Si0₂ at 350 °C under a high pressure of oxygen, contains [Ag₆]⁴⁺ clusters (average oxidation state 2/3) and should be formulated at [Ag₆]⁴⁺ (Ag⁺)₄ (SiO₄ ^{4 -})₂.

Ag and Au forms numerous cluster compounds in which the metals are zero valent or subvalent. Examples are Ag₆ Fe₃ (CO)₁₂ {HC(PPh₂)₃} and paramagnetic cluster ions [Ag₁₃ Fe₈ (CO)₃ ²]^4-.