Protoplast isolation and culture (1)
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2. Prokaryotic and eukaryotic cell structure
Prokaryotic and Eukaryotic cell
All living cells can be classified as
Prokaryotes cells: pre-nucleus
the Greek words pro (before) and karyon (nucleus).
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All living cells can be classified as
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science focusing on the structure,
function, evolution, mapping, and editing
of genomes. A genome is an organism's
complete set of DNA, including all of its
genes. In contrast to genetics, which
refers to the study of individual genes and
their roles in inheritance, genomics aims
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quantification of genes, which direct the
production of proteins with the assistance
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1980 Nobel Prize in chemistry for independently
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Prokaryotic and eukaryotic cells differ structurally. Prokaryotic cells lack membrane-bound organelles and have no nucleus, while eukaryotic cells have organelles like mitochondria and a nucleus enclosed in membranes. Both cell types share chemical and metabolic similarities like genetic material, cell membranes, and similar metabolic reactions, but prokaryotes are typically smaller without internal compartments.
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Protoplast isolation and immobiliz by Dr.U.Srinivasa
The document discusses protoplasts, which are plant cells that have had their cell walls removed, leaving the cell membrane and organelles. It describes methods for isolating protoplasts from plant tissues using either mechanical or enzymatic methods. The enzymatic method uses enzymes like pectinase and cellulase to break down the cell wall. Protoplasts have various applications including isolating cell organelles and studying cell structures. The document also discusses immobilizing enzymes by binding them to inert matrices, which has benefits like reusability and stability. Methods of immobilization include adsorption, covalent binding, and entrapment in gels.
Isolation of protoplast in plant tissue culture.
The document discusses the isolation of protoplasts from plant cells. There are two main methods for isolating protoplasts - mechanical and enzymatic. The enzymatic method uses enzymes to digest the cell wall and is more widely used as it works for a variety of plant tissues and causes less damage to the cells. Key steps in the enzymatic method include incubating plant tissue in enzyme solutions, filtering to separate protoplasts from debris, and centrifugation to purify the protoplasts. Isolated protoplasts can be used for cell fusion between unrelated plant species and for genetic modification of plants. Factors like plant species, age of donor tissue, and pre-treatment of tissue affect the viability
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Prokaryotes vs eukaryotes
The document compares and contrasts prokaryotic and eukaryotic cells. Prokaryotes lack a nucleus and membrane-bound organelles, while eukaryotes have a nucleus enclosed in a nuclear envelope and various membrane-bound organelles. Some key differences include prokaryotes being typically smaller (1-10 μm) than eukaryotes (10-100 μm), having circular DNA versus eukaryotic linear DNA, and dividing via binary fission rather than mitosis and meiosis. Prokaryotic cells also generally lack internal membranes, mitochondria, chloroplasts and a complex cytoskeleton.
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2. The program covers topics such as the diversity of invertebrate and vertebrate animals, cell biology, genetics, physiology, ecology, and applied areas like biotechnology.
3. Courses involve both theory and practical components, with the later semesters focusing more on advanced topics and including projects.
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Prokaryotes are single-celled organisms without membrane-bound organelles that include bacteria and archaea. They have distinct characteristics like DNA that is not enclosed in a nucleus, lack of histones, and cell sizes typically between 1-5 micrometers. Prokaryotes reproduce through binary fission and have various internal structures like ribosomes, cytoplasmic membrane, and nucleoid containing DNA. They also have external structures such as flagella, pili, and capsules that provide motility and attachment abilities. The cell wall and plasma membrane serve as protective barriers around the cell.
Bacteria Project
This document summarizes a study conducted by researchers at the University of Puerto Rico at Cayey to identify soil bacteria capable of producing novel antibiotics. Soil samples were collected from two sites and diluted to isolate individual bacterial colonies. Colonies were purified, stained, and had their DNA analyzed. The isolated bacteria were tested for antibiotic resistance and their ability to inhibit the growth of other bacteria, which could indicate antibiotic production. The goal was to find bacteria producing compounds similar to teixobactin, a potent antibiotic discovered from uncultured soil bacteria.
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This document discusses the key differences between prokaryotic and eukaryotic cell structure and function. It notes that while both have the same basic biological molecules, eukaryotes have membrane-bound organelles that compartmentalize processes. Prokaryotes like bacteria are smaller and lack a nucleus or organelles. They have distinctive cell walls and internal structures like circular DNA, ribosomes, and may form endospores. The document outlines the structural features of prokaryotic cells.
Cell
Cells are the basic unit of life, and all living things are made up of one or more cells. Cells contain cytoplasm enclosed within a membrane and include organelles and biomolecules. Cells can be either unicellular, like bacteria, or multicellular, like plants and animals. The first person to observe cells was Anton van Leeuwenhoek in the 1600s, and later researchers discovered cell structures like the nucleus. In the 1830s, scientists proposed the cell theory - that cells are the fundamental unit of life and all cells come from pre-existing cells. Cells are classified as prokaryotic or eukaryotic, with eukaryotic cells having membrane-bound organelles and a nucleus.
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This document provides definitions and details about cell biology. It begins by defining cell biology and its history. It describes the origin of life and cells, including theories about abiogenesis and the endosymbiotic theory of the origin of eukaryotic cells. It discusses cell theory and the two main types of cells - prokaryotic and eukaryotic cells. Further, it provides information about key cellular structures like the cell membrane, nucleus, and their structure and functions. The document is a comprehensive overview of foundational concepts in cell biology.
Prokaryotic cell
Prokaryotic cells lack membrane-bound organelles and have a circular chromosome located in an irregular region called the nucleoid. They may contain extrachromosomal DNA in plasmids. Prokaryotic cells share components like the plasma membrane, ribosomes, and DNA. Some prokaryotes perform photosynthesis using thylakoid or chromatophore membranes. Prokaryotic cells may also contain mesosomes, cell walls, glycocalyces, S-layers, and filamentous appendages like flagella, fimbriae, and pili that allow interaction with the environment.
Prokaryotic cell
Prokaryotes are unicellular organisms that lack internal membrane-bound structures. They are divided into bacteria and archaea. Prokaryotic cells lack a nucleus but contain a nucleoid region where DNA is located. Archaeal membranes contain isoprene lipids instead of fatty acids found in bacteria. Prokaryotes reproduce through binary fission and exchange genetic material through transformation, transduction, and conjugation.
Bacteria and viruses
Bacteria and viruses differ in their structure. Bacteria have cell walls, lack organelles, and divide through binary fission. They can be gram-positive or gram-negative. Viruses are much smaller, lack cells, and contain genetic material surrounded by a protein coat. They invade host cells and use the cell's machinery to replicate. Microorganisms play important roles in decomposing organic matter and recycling carbon through the environment.
THE PROKARYOTIC CELL
Prokaryotic cells include bacteria, blue-green algae, mycoplasma, and PPLO. They lack a nucleus and other membrane-bound organelles. The cell envelope consists of an outer glycocalyx layer, a peptidoglycan cell wall, and an inner plasma membrane. The cell contains genomic DNA and sometimes plasmid DNA. Ribosomes are the site of protein synthesis and prokaryotic ribosomes are 70S. Inclusion bodies store reserved materials in the cytoplasm.
eukaryotic n prokaryotic cells
The document discusses the key differences between prokaryotic and eukaryotic cells. Prokaryotic cells lack a nucleus and membrane-bound organelles, while eukaryotic cells have a nucleus enclosed in a nuclear membrane and membrane-bound organelles. The genetic material in prokaryotes is a single loop of DNA, whereas in eukaryotes it is organized into linear chromosomes within the nucleus. Prokaryotes tend to be unicellular while eukaryotes can be unicellular or multicellular. The document then provides more details on the structures and components of prokaryotic and eukaryotic cells.
Bacterial morphology & anatomy
This document discusses the morphology and anatomy of bacterial cells. It describes prokaryotic cells as being much smaller and simpler than eukaryotic cells, lacking membrane-bound organelles. The typical bacterial cell has a cell wall, cell membrane, cytoplasm, and sometimes additional structures like flagella or spores. The cell wall provides shape and rigidity, and its chemical composition divides bacteria into Gram-positive and Gram-negative types. Bacterial cells range in size from 0.2 to 1.5 micrometers and can have different shapes.
Chap 2 fungal cell
The fungal cell wall is composed of various components that give shape, strength, and protection to fungi. The main components are chitin, glucan, and proteins. Chitin is a nitrogenous polysaccharide made of N-acetylglucosamine that forms microfibrils providing structure. Glucan is a polysaccharide of glucose monomers linked by beta glycosidic bonds that is a major component of fungal cell walls. Glycoproteins and proteins are also embedded in the cell wall matrix and contain carbohydrates. Together these components form the primary wall and secondary wall layers that define fungal cells.
prokaryotic cell
Prokaryotic cells are the earliest and most primitive forms of life on Earth. They include bacteria and archaea and some are capable of photosynthesis. Prokaryotic cells have no nucleus and their DNA is located in the nucleoid region of the cytoplasm. They reproduce through binary fission and can also undergo genetic recombination through conjugation, transformation, or transduction to generate variation. Prokaryotic cells play important roles in various environments and as normal flora in humans and animals.
Introduction to basic concept of genomics
Define the following and state the techniques used for the study of the following_a) Genomics b) Proteomics c) Transcriptomics d) Metabolomics e) Phenomics
Genomics is an interdisciplinary field of
science focusing on the structure,
function, evolution, mapping, and editing
of genomes. A genome is an organism's
complete set of DNA, including all of its
genes. In contrast to genetics, which
refers to the study of individual genes and
their roles in inheritance, genomics aims
at the collective characterization and
quantification of genes, which direct the
production of proteins with the assistance
of enzymes and messenger molecules. Proteomics involves the study of the structure of all proteins encoded in a fully sequenced genome which is also referred as proteomos.
Frederick Sanger and Walter Gilbert shared half of the
1980 Nobel Prize in chemistry for independently
developing methods for the sequencing of DNA.
B.sc. (micro) i em unit 3.1 bactria
This document provides an overview of bacterial morphology and classification. It defines prokaryotic cells and describes their main structures, including the cell wall, plasma membrane, cytoplasm, and additional organelles like flagella and spores. It explains that the chemical composition of the cell wall forms the basis for classifying bacteria as either Gram-positive or Gram-negative. The key differences between prokaryotic and eukaryotic cells are also summarized in tables.
Prokaryotes _Eukaryotes
Prokaryotic and eukaryotic cells differ structurally. Prokaryotic cells lack membrane-bound organelles and have no nucleus, while eukaryotic cells have organelles like mitochondria and a nucleus enclosed in membranes. Both cell types share chemical and metabolic similarities like genetic material, cell membranes, and similar metabolic reactions, but prokaryotes are typically smaller without internal compartments.
Bacteria lec
Microorganisms can be classified into three kingdoms: plants, animals, and protists. The kingdom protists includes unicellular microorganisms like bacteria, fungi, protozoa, and algae. Bacteria and archaebacteria are types of prokaryotes, while fungi, algae, protozoa and slime molds are eukaryotes. Bacteria are very small, typically measuring between 2-5 micrometers, and require microscopy to be studied. Bacteria have different shapes including cocci, bacilli, coccobacilli, and spirilla. The bacterial cell envelope includes a cell wall and plasma membrane that protects the cell and gives it structure and shape.
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Protoplast isolation and immobiliz by Dr.U.Srinivasa
The document discusses protoplasts, which are plant cells that have had their cell walls removed, leaving the cell membrane and organelles. It describes methods for isolating protoplasts from plant tissues using either mechanical or enzymatic methods. The enzymatic method uses enzymes like pectinase and cellulase to break down the cell wall. Protoplasts have various applications including isolating cell organelles and studying cell structures. The document also discusses immobilizing enzymes by binding them to inert matrices, which has benefits like reusability and stability. Methods of immobilization include adsorption, covalent binding, and entrapment in gels.
Isolation of protoplast in plant tissue culture.
The document discusses the isolation of protoplasts from plant cells. There are two main methods for isolating protoplasts - mechanical and enzymatic. The enzymatic method uses enzymes to digest the cell wall and is more widely used as it works for a variety of plant tissues and causes less damage to the cells. Key steps in the enzymatic method include incubating plant tissue in enzyme solutions, filtering to separate protoplasts from debris, and centrifugation to purify the protoplasts. Isolated protoplasts can be used for cell fusion between unrelated plant species and for genetic modification of plants. Factors like plant species, age of donor tissue, and pre-treatment of tissue affect the viability
Protoplast culture
Protoplasts are naked plant cells without the cell wall, but they possess plasma membrane and all other cellular components. They represent the functional plant cells but for the lack of the barrier, cell wall. Protoplasts of different species can be fused to generate a hybrid and this process is referred to as somatic hybridization (or protoplast fusion). Cybridization is the phenomenon of fusion of a normal protoplast with an enucleated (without nucleus) protoplast that results in the formation of a cybrid or cytoplast (cytoplasmic hybrids).
PROTOPAST ISOATION, PROTOPAST FUSION AND SOMATIC HYBRIDISATION.pptx
Protoplasts are plant cells that have had their cell walls removed, leaving the plasma membrane as the outer layer. They can be isolated from various plant tissues like leaves and cell suspension cultures through enzymatic digestion of the cell walls using enzymes like cellulases, pectinases, and hemicellulases. There are two main methods for isolating protoplasts - sequential and mixed enzymatic methods. In the sequential method, pectinase treatment is followed by cellulase treatment, while the mixed method uses a combination of enzymes simultaneously. After enzymatic treatment, protoplasts are collected, washed, and cultured on appropriate media to induce cell division and regeneration of plants.
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This document summarizes the structures of prokaryotic bacterial cells. It describes structures external to the cell wall like capsules, slime layers, S layers, pili, and flagella. It also describes the cell wall, cell membrane, cytoplasm, ribosomes, mesosomes, nucleoid, and inclusion bodies that make up the internal structures of bacterial cells. Each structure is defined and its composition and functions are explained.
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This document summarizes methods for isolating and culturing plant protoplasts. Protoplasts are plant cells that have had their cell walls removed. The document describes two methods for isolating protoplasts - mechanical and enzymatic. It also discusses purifying the protoplasts, assessing their viability, culturing them in liquid media to regenerate cell walls, and the potential applications of protoplast culture and regeneration including plant breeding techniques.
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This document provides an introduction and history of plant tissue culture. It discusses various types of plant tissue cultures including callus culture, cell suspension culture, protoplast culture, meristem culture, anther culture, somatic embryogenesis, and ovary/ovule culture. The key applications of these culture techniques are the production of virus-free plants, mass production of desirable genotypes, production of haploid plants, and genetic transformation.
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This presentation slide is helpful for undergraduate and postgraduate student of biotechnology, plant science and botany student.
In this video I am describe protoplast isolation method.
Protoplast isolation,culture & fusion
The isolation, culture and fusion of protoplasts is a fascinating field in plant research. Protoplast isolation and their cultures provide millions of single cells (comparable to microbial cells) for a variety of studies.
Somatic hybridization
The document discusses somatic hybridization through the fusion of protoplasts from different plant species. It describes:
1. The process of somatic hybridization which involves isolating protoplasts from plant tissues, fusing the protoplasts from different species using chemical or electrical methods, selecting hybrid cells, culturing the hybrid cells and regenerating hybrid plants.
2. Methods for isolating viable protoplasts including enzymatic and mechanical methods. Enzymatic isolation uses cellulase, hemicellulase and pectinase enzymes.
3. Techniques for purifying isolated protoplasts such as filtration, centrifugation, flotation and density buffer methods to remove
Protoplast culture
This document summarizes the process of isolating plant protoplast cells. It defines a protoplast as a plant cell without its cell wall. There are two main methods for isolating protoplasts - mechanical and enzymatic. The enzymatic method uses enzymes like pectinase and cellulase to gently dissolve the cell wall and is preferred. The steps of the enzymatic isolation process are described, including incubating tissue in the enzyme solution, filtering, washing, and testing viability. Finally, some applications of isolated protoplasts in research are mentioned like transformation and somatic hybridization.
Protoplast Culture
This document summarizes the process of isolating plant protoplast cells. It defines a protoplast as a plant cell without its cell wall. There are two main methods for isolating protoplasts - mechanical and enzymatic. The enzymatic method uses enzymes like pectinase and cellulase to gently dissolve the cell wall and is preferred. The steps of the enzymatic isolation process are described, including incubating tissue in the enzyme solution, filtering, washing, and testing viability. Finally, some applications of isolated protoplasts in research are mentioned like transformation and somatic hybridization.
Unit 2 cells
The document discusses the history and development of microscopy and cell theory. It begins with early microscopes like hand lenses and single lens microscopes used to first observe cells. Today, electron microscopes like SEM and TEM are used. Key contributors included Hooke, who first observed cells, Leeuwenhoek who discovered bacteria and cells, and Schwann and Schleiden who developed the original cell theory. The document then describes plant and animal cell structures like cell walls, chloroplasts and organelles in detail. It explains membrane structure and transport mechanisms. The importance of microscopy in observing cells is also highlighted.
Synthetic cells
SYNTHETIC CELLS
An artificial cell or minimal cell or synthetic cell is an engineered particle that mimics one or many functions of a biological cell.
Artificial cells are biological or polymeric membranes which enclose biologically active materials.
A "living" artificial cell has been defined as a completely synthetically made cell that can capture energy, maintain ion gradients, contain macromolecules as well as store information and have the ability to mutate.
DEFINITION
EXAMPLE
SYNTHETIC BIOLOGY
Synthetic biology is a multidisciplinary area of research that seeks to create new biological parts, devices, and systems, or to redesign systems that are already found in nature.
Due to more powerful genetic engineering capabilities and decreased DNA synthesis and sequencing costs, the field of synthetic biology is rapidly growing
HISTORY
BOTTOM-UP APPROACH FOR CONSTRUCTING SYNTHETIC CELLS
A bottom-up approach is commonly used to design and construct genetic circuits by piecing together functional modules that are capable of reprogramming cells with novel behavior.
CELL ENCAPSULATION METHOD
Cell microencapsulation technology involves immobilization of the cells within a polymeric semi-permeable membrane that permits the bidirectional diffusion of molecules such as the influx of oxygen, nutrients, growth factors etc. essential for cell metabolism and the outward diffusion of waste products and therapeutic proteins.
TECHNIQUES USED FOR THE PREPARATION OF EMULSION
1- high pressure homogenization
2- microfluidization
3- drop method
4- emulsion method
MEMBRANES OF SYNTHETIC CELLS
THE MINIMAL CELL
A minimal cell is one whose genome only encodes the minimal set of genes necessary for the cell to survive.
THE SYNTHETIC BLOOD CELLS
Synthetic red blood cells mimic natural ones, and have new abilities
APPLICATIONS OF SYNTHETIC CELLS
1- DRUG RELEASE AND DELIEVERY
2- GENE THERAPY
3- ENZYME THERAPY
4- HEMOPERFUSION
5- OTHER APPLICATIONS
FUTURE OF SYNTHETIC CELLS AND BIOLOGY
ACHIEVEMENTS
HEALTH AND SAFETY ISSUES
ETHICS AND CONTROVERSIES
REFERENCES
THANK YOU
3. prokaryotic vs eukaryotic cells
This document discusses and compares prokaryotic and eukaryotic cells. It provides information on their key structures and functions. Prokaryotic cells are single-celled organisms that lack a nucleus and include bacteria. They have a cell membrane, cell wall, cytoplasm, ribosomes, and can reproduce asexually. Eukaryotic cells are larger, have membrane-bound organelles and a true nucleus, and can reproduce both sexually and asexually. Examples of eukaryotes include plants, animals and fungi. The document provides details on specific structures like the nucleus, cell membrane, and flagella in both cell types.
Somatic ybridization and its application
This document discusses somatic hybridization, which involves fusing plant protoplasts from two different species or varieties to create a hybrid plant. It describes the process of somatic hybridization, including isolating protoplasts, fusing them using spontaneous or induced methods, selecting hybrid cells, and regenerating plants from hybrid callus tissue. The advantages are producing novel hybrids and transferring genes between incompatible species. The limitations include low regeneration rates and viability of fused cells. Somatic hybridization has applications in crop improvement by introducing traits like disease resistance from wild relatives.
Biologi grade7
The document discusses the key characteristics of fungi, plants, and animal cells. It provides details on the structures and organelles found within these eukaryotic cell types, including their cell walls, nuclei, mitochondria, chloroplasts, and means of reproduction. Major differences highlighted include fungi having cell walls containing chitin, plants having cell walls containing cellulose and chloroplasts for photosynthesis, and animal cells lacking cell walls.
PROTOPLAST ISOLATION & CULTURING TECHNIQUES.
All about what is protoplast, how to isolate protoplast & technques to culture them.
Also the potent applications & somatic hybridization.
Lesson 4 bio101 (c)Dr. Evangelista
The protoplast is composed of the nucleus and cytoplasm, which contains membrane-bound organelles. The cytoplasm includes the cytosol and endomembrane system. The endomembrane system is made up of the nuclear envelope, endoplasmic reticulum, Golgi apparatus, vacuoles, and plasma membrane. It works to compartmentalize cellular processes. The protoplast also contains cytoskeletal elements like microtubules, microfilaments, and intermediate filaments that provide structure and enable cellular movement. Ribosomes are found throughout the cytoplasm and carry out protein synthesis.
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Protoplast isolation and culture (1)
- 1. Submitted by , R. Yamuna, 2nd M.Sc. Biotechnology, Annamalai University. PROTOPLAST ISOLATION AND CULTURE
- 2. SYNOPSIS i. What is a protoplast? ii. What are the methods used ? iii. Enzymatic method. iv. Reagents and their role. v. Procedure . vi. Protoplast culture.
- 3. WHAT IS A PROTOPLAST ? Living part of the plant cell. With cytoplasm and nucleus. No cell wall. Isolated from – roots , leaves, tubers, root nodules, endosperm. Convinent method – leaf. Reason – mesophyll cells loosely arranged.
- 4. WHAT ARE THE METHODS USED ? Mechanical method. Enzymatic method. 1.MECHANICAL METHOD – cutting / breaking the cell wall. 2.ENZYMATIC METHOD – digestion of cell wall with enzymes.
- 5. ENZYMATIC METHOD Leaf sterlization, removal of epidermis. Plasmolyse d cells Release of isolated cells. Isolated protoplasm Plasmolyse d cells NON SEQUNTIAL SEQUNTIAL Protoplasm released. Protoplasm released Pectinase + cellulase Pectinase
- 6. REAGENTS AND THEIR ROLE 70 % Ethanol – sterlization and washing 70% ethanol+30%water – easily penetrate into cell . More polar – better osmosis. 2% Sodium Hypochloride – disinfectant / antimicrobial agent. Mannitol – plasmolysis , hyperosmotic environment . ( osmoticum )
- 7. Pectinase – breakdown protein – holding cells together . Cellulase – digest cellulose. Sorbitol – plasmolysis , hyperosmotic environment (osmoticum). Sucrose – density gradient – centrifugation – interface.
- 8. (cont.) Potassium dextran sulphate – release DNA from DNA histone, inhibits binding of RNA to ribosomes. Distilled water. Centrifuge. Micro pipittes. Petri plates.
- 9. PROCEDURE Surface sterlize (70%) and Wash – 2% Na hypochloride Peel the lower epidermis – place in petri dish – pectinase Agitate – slowly . After 15- 20 minutes – replace the enzyme mixture 0.5% macerozyme,0.3% potassium dextran sulphate in 13% mannitol
- 10. (cont.) Incubate – 1 hr. Filter the isolated cells – nylon membrane Isolated cells+enzyme mixture Centrifuge 100xg -1m Centrifuge 100xg -1m Decant the supernatant and wash the pellet with mannitol
- 11. Clean the protoplast – 25% sucrose Sucrose+pellet+0 .4sorbitol+10mm cacl2. Sample - interface Sample collected . Added to salt solution – 0.2%w/v cacl2 and 2.5% Hcl PROTOPLAST Centrifuge at low speed
- 12. PROTOPLAST CULTURE Nagata and takebe medium CULTURE TECHNIQUES
- 13. THANK YOU