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Basics of Biotechnology

Basics of Biotechnology

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Basics of Biotechnology

BIOTECHNOLOGY

Starch Slurry in Water

(Liquefaction Bacterial amylase, 550C +Dextrins)

(Saccarification Fungal amylase)

Maltose

(Amylo glucosidae)

Glucose

(Partial isomerization)

Fructose + glucose

· Variation created in tissue culture is called “Somaclonal variation”.

· The specialized cells (B – cells) in the spleen and lymph glands release antibodies.

· Monoclonal antibodies is applied to a group of identical antibodies recognizing the same antigen.

· Callus – A tissue arising from disorganized proliferation of cells.

· Subculture – A septic transfer of a part of a culture to fresh medium.

· Passage time – The time interval between two successive sub cultures.

· Suspension culture – Cells (or) cell aggregates culture in liquid medium.

· Batch culture – Cell suspension grown in a fixed volume of liquid medium.

· Explant – A plant organ (or) piece of tissue used to initiate culture.

· Excise – To remove a piece of tissue (or) an organ from its parent source.

· Meristemoid – Cluster of meristematic cells with in Mcallus with a potential to form primordia.

· Embryoid – Non zygote embryo formed in culture.

· In vitro – “In glass”; In vivo – “In life”.

· Parasexual hybridization (or) somatic hybridization – Hybridisation by non sexual methods.
Eg: Potoplast fusion.

· Heterokaryon – A cell in which 2 (or) more nuclei of unlike genetic make up are present.

· Synkaryon – Hybrid cell produced by the fusion of nuclei in homokaryon.

Bits: In mass selection genotype of selected plants is not known since progeny test is not done.

· During plant introduction the introduced material should be accompanied by a “Phytosanitory” certificate.

· Allotctraploid otherwise termed as “Amphidiploid”.

· The homozygous lines developed by controlled selfing in cross pollinated crops are called “Inbred lines”.

· Parent to progeny relationship is known in Pedigree method.

· The difference between the mean of the original population and mean of the selected plants is termed as “Selection differential”.

· Mass selection is based on “Phenotype”.

· The percentage of plants selected from a population to be advanced to next generation is called “Selection intensity”.

· The performance of a double cross hybrid in maize can be predicted based on the performance of non parental single crosses.

· Cytoplasmic male sterility is used for production of vegetables, and ornamental plants.

· Genetic male sterility is used for hybrid production of “Castor”.

· Modified flower – Bulbils.

· The first cytoplasmic genetic male sterile line in Sorghum – CSH -7.

· Maximum amount of segregations are observed in F2 generation.

· Bajra composite – Balaji, Maize synthetic – Amber.

Leaf blight resistant cotton – Varalakshmi.

· Induced mutant in Groundnut MC -4, Resistance to tikka leaf spot – Ah – 45.

· Heteroplast – Cell containing foreign organells.

· Genetic engineering – manipulation of the genetic archilecture of the organism at DNA level (or) molecular level.

Explant——–> Callus——-> Plant let 

Didifferentiation     Redifferentiation

· Tissue culture techniques are being used for effective vegetative propagation of horticultural (or) agricultural crops. These techniques are called “Micropropagation”.

· Activated charcoal stimulates embryogenesis in plants.

· The most popular medium for anther culture is N6 medium.

· In anther culture in rice pretreatment is done with “ethereal”.

· Iron is crucial for pollen embryo development in tobacco.

· Dry heat – metals, glassware, wet heat – paper material, liquids.

· Some media components are unstable at high temperature and must be sterilized by “Ultra filtration” at high temperature.

· Working area is generally surface sterilized with either ethanol (or) isopropanol.

· Organogenesis in tissue culture is controlled mostly by a balance between cytokinin and auxin.

· In callus of Medicago sativa high kinetin, 2,4 – D ratio stimulates “rhizogenesis” high 2,4 – D; Kinetin ratio – caulogenesis.

· For embryo culture MS medium and Monnier’s medium favorable.

· ‘Sucrose’ is added to embroyo culture to maintain suitable osmatic pressure.

· “Casein – hydrolysate” (CH) amino acid complex increases embryonic differentiation and the size of the embryo.

· Coconut milk is referred to as “Endosperm factor”.

Most of the responding sps to endosperm culture – Euphorbiaceae, Santalaceae, Loranthaceae.

· Cultured endosperm of coffee does synthesize – ‘Caffeine’.

· The middle lamella is dissolved by using “Pectinase”.

· Protoplast may be cultured either by “liquid drop method” (or) plating method (Common method).

· To induce protoplast fusion suitable chemical agents called “Fusogen” are used. These include PEG, Polyvinyl alcohol, NaNO3.

· Cybrid may be defined as plant (or) cell which is a cytoplasmic hybrid produced by fusion of protoplast and Lytoplast.

· Cybrids have two different cytoplasm and one nuclei.

· Vitrification – In repeated cycles of invitro shoot multiplications often a percentage of cultures show water soaked, almost translucent leaves. Such inhibits a rate of growth.

· The heart of the biotechnology is the “bioreactor” where the bio-transformation takes place.

· Fluid flow characteristics obey Newton laws of viscosity.

· The basic types of fermentation processes – Semisolid fermentation and submerged fermentation.

· “Enzyme assay” is to determine how much the enzyme of known characteristic is present in the tissue homogenate, fluid (or) partially purified extract.

· Enzymes purification follows an important step called “Extraction”

· The crude extract is treated with absorbant gels like “Zn(OH)2 for removal of pigments.

· “Affinity Chromatography” is a very bio specific one and it is ideally suited for the separation of one protein from all other.

· “Urakinase” from human urine, used for cleaning wounds and removal of blood clots.

· Nutrients for cancer cells – A sparagine, glutamine are removed by asparaginase and glutaminase.

· Glucose oxidase for determining blood glucose.

· LDH (Lactic dehydrogenase) for lactate and pyruvate.

· A glucose oxidae – Catalase is effective in removing O2.

· Some of the soil enzymes – Cellulase, nitrogenases, urease etc.

· Biosensors can measure sugars, proteins, hormones in body fluids pollutant in water.

· Enzyme biosensors – Alcohol Oxidase, Invertase, glucose oxidase,
Microbial bio sensors – Brevibacterium incto fermentum.

· Recombinant DNA is method of transferring relatively small amount of genetic information from one cell type to the other.

· Enzymes which synthesize DNA – Terminal transferase, DNA polymerase Restriction endonuclease.
· Enzymes that degrade DNA – Nuclease S1, Exonuclease III.
Restriction Endonuclease.

· Enzymes that joins DNA sequence – T4 DNA ligase.

· Enzymes which modify the 5 terminal of DNA – Calf intestinal
Phosphatase, T4 polynucleotide kinase.

· The sequence recognized by restriction enzymes are “Palindromes”.

· In case of mammalian cells, there is only one potential vector the “Simian virus” (SV –40).

· Many animals “tumor virus” contain reverse transcriptase.

· A useful plasmid is “pBR 322” (resistance to tetracycline and ampicillin) it has 4322 bp.
Transgenic organisms – These are the outcome of the recombinant DNA technology.

· Cauliflower mosaic virus (CMV) and Gemini virus have also been used as vectors for plant transformations.

· Yeast are the most popular eukaryotic cell for cloning.

· Bacteria involved in fermentation processes are “Chemo-organotrophes”.

· The first therapeutic agent produced by recombinant DNA technology using E. coli is “human insulin”.

· E. coli contain endotoxins and pyrogenic lipopoly saccharides.

· Hybridoma cells are constructed by fusing a mouse myeloma and B. Cells (skin cancer cells).

· Starch is the important – CHO currently used in fermentation process.

· Batch and fed batch fermentation – Closed system.

· All the substrate is added at the begining of fermentation in batch but in fed batch they are added in increments throughout the process.

· Continuous fermentation – Open system in which medium is continuously added.

· Stock cultures are kept in the laboratory in freeze dried from deep frozen in liquid nitrogen.

· Some of the flocculating agents – Isin glass (a collagen type protein derived from a tropical fish) gelatin, tannic acid.

· Centrifugation may be the only practicable separation of cells from viscous media.

· Tangential flow filtration (or) cross flow filtration is an effective method for separation of cells from liquid where high value products are involved.

· Citric acids and lactic acids produced by fermentation are precipitated as their “Calcium salts”.

· The most commonly used ion-exchange celluloses are “Diethyl amino ethyl” (DEAE) and carboxy methyl, cellulose anion and cation exchanges respectively.

· The most commonly studied secondary metabolites are “Antibiotics”.

· For gelling more amount of starter culture, we can used “Seed rank fermentor”.

Bits: –
· The dis advantages with micropropagation of adult tree is browning of the medium.

· Somaclonal variation is the consequence of genetic abnormalities.

· Proloplast obtained from endosperm cells gave best results in tobacco.

· The capacity of a cell to give rise to a whole plant – “totipotency”.

· The artificial seed of carrot somatic embryo’s are coated by polyethelene oxide. (PEO).

· The filamentous fungus that is genetically engineered for production of proteins of mammalian origin is Aspergillus niger.

· The important yeast used in industry – Saccharomyces cerviciae.

· S.I unit of enzyme – “Ketal”.

· Cosmids vectors are used for cloning large fragments of DNA.

· Detergents – Proteases.

· In gel chromatography molecular wt. is the criteria for separation.

· For production of ‘SCP’ Protein: RNA ratio must be highest

· The amount of enzyme per unit wt. of protein is termed as “Turn over of the enzyme”.

· Enucleate protoplast is termed as cytoplast.

· Synthetic seeds – Somatic embryo genesis, virus free plants – shoot tip culture.
Somatic hybrids – Protoplast fusion.

· Inter specific hybrids – Embryo culture – Rare hybrids also
Inter generic hybrids – Protoplast culture.
Triploids – Endosperm culture.
Uniform diploids – Meristem culture.

· A vehicle which is used to carry other species of DNA is called vector.

· Plant oils used in industrial fermentation media are palm oil

· Human viral protein produced on large scale for medical purpose – Interferon.

· Organism used for riboflavin (B2) fermentation Ashbyo gossypii.

· Procem of origin of shoot, bud (or) roots from tissue (or) suspension culture organiogenesis.

· Cells (or) cell aggregates culture in liquid medium is termed as suspension culture.

· The most serious objection for clonal propagation through callusing is due to genetic instability.

· Usual source for large scale production of enzymes is Microbial cells.

· Homozygous tobacco plant developed through anther culture –F.

· The pinocytosis property of protoplasts is utilized in transfer of genome of the one species to others.

· Small circular DNA found inside some bacteria is Plasmid.


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