This document discusses molecular farming, which uses genetic engineering techniques to produce commercial products in plants. Potential vehicles for protein production include the leaves of tobacco and alfalfa for bulk production and crops like tomato, potato, maize, wheat and rice for edible products. Stable and transient expression strategies can be used. Examples of molecular farming applications include producing antibodies and vaccines in plants, as well as metabolic engineering of carbohydrates like polyfructans and lipids like polyhydroxyalkanoates for bioplastics. Recombinant proteins like hirudin have also been produced in transgenic plants.
This document discusses molecular farming, which uses genetic engineering techniques to produce commercial products in plants. Potential vehicles for protein production include the leaves of tobacco and alfalfa for bulk production and crops like tomato, potato, maize, wheat and rice for edible products. Stable and transient expression strategies can be used. Examples of molecular farming applications include producing antibodies and vaccines in plants, as well as metabolic engineering of carbohydrates like polyfructans and lipids like polyhydroxyalkanoates for bioplastics. Recombinant proteins like hirudin have also been produced in transgenic plants.
This document discusses molecular farming, which uses genetic engineering techniques to produce commercial products in plants. Potential vehicles for protein production include the leaves of tobacco and alfalfa for bulk production and crops like tomato, potato, maize, wheat and rice for edible products. Stable and transient expression strategies can be used. Examples of molecular farming applications include producing antibodies and vaccines in plants, as well as metabolic engineering of carbohydrates like polyfructans and lipids like polyhydroxyalkanoates for bioplastics. Recombinant proteins like hirudin have also been produced in transgenic plants.
This document discusses molecular farming, which uses genetic engineering techniques to produce commercial products in plants. Potential vehicles for protein production include the leaves of tobacco and alfalfa for bulk production and crops like tomato, potato, maize, wheat and rice for edible products. Stable and transient expression strategies can be used. Examples of molecular farming applications include producing antibodies and vaccines in plants, as well as metabolic engineering of carbohydrates like polyfructans and lipids like polyhydroxyalkanoates for bioplastics. Recombinant proteins like hirudin have also been produced in transgenic plants.
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Molecular Farming in Plants
Dr. Neelu Joshi
Assistant Professor Department of Biotechnology and Bioinformatics Padmashree Dr. D.Y. Patil University Navi Mumbai
Applications of r DNA technology in Agriculture
Crop protection against various biotic and abiotic
stresses Modification of production traits /introducing novel traits in crops Improving colour and smell of flower, taste of food crops Modifying starch, protein and oil content Facilitating the accumulation of vitamins and minerals in food Altering the properties of fruits and vegetable Introducing male sterility Increasing the levels of total soluble solids Metabolic engineering Molecular farming
Molecular Farming Application of genetic engineering techniques to
the synthesis of commercial products in plants.
It includes:
-enhanced production of products that are
already extracted from plants. -manufacture of compounds novel to plants.
Molecular farming in plants Potential vehicles for protein production: for bulk production- leaves of tobacco &alfalfa, for edible products- tomato, potato, maize, wheat, rice Production strategies Stable expression: constitutive and tissue specific Transient expression: using plant virus (Cp gene)as vector
Molecular farming for carbohydrate
Polyfructans (G-F-Fn) Soluble polymer of fructose, synthesized and stored in vacuole Fructans are used as carbohydrate reserves Ex. Inulins-major storage carbohydrate found in onion bulbs Levans-leaves and stem of grasses Glucose-fructose-(fructose)n i.e. G-F-Fn Biosynthetic pathway of fructans 1. G-F+G-F = G-F-F+G SST enzyme 2. G-F-Fn+G-F-F = G-F-Fn+1 +G-F FFT enzyme. Sucrose is the initial acceptor molecule of the chain, so the first sugar in the fructan chain is always glucose. However, for each remaining fructose residue added to the chain a glucose residue is released, which in plants is transported back out of the vacuole into the cytosol. sacB gene(produces levansucrase in one step) of B. subtilis +vacuolar targeting sequence from yeast, in tobacco. Sac B (B.amyloliquifaciencs) +zein promoter +vacuolar targetting seq. expressed in maize endosperm
Fructanbeet- transgenic sugarbeet Production of short oligofructans GF2, GF3, GF4. Gene for enzyme 1-SST from Jerusalem artichoke Applications: 1.In neutraceutical market 2.As substitute of sugar in foods and drinks 3.Low-calorie sweetner 4.Encourage growth of beneficial gut flora and
cause acidification of the digestive tract, which
inhibits E.coli.
Molecular farming for lipids Polyhydroxyalkanoates (PHAs) form bioplastics
Bioplastics Currently produced by microbial fermentation Polyhydroxybutyrate (PHB)-best characterized PHA found as
intracellular inclusions in bacteria e.g. Alcaligenes eutrophus.
Biosynthetic pathway in A.c. Pha B Pha Acetyl-CoA Pha acetoacetyl coA 3-hydroxybutyrl-coA c A PHB Cytoplasm plastid Pha Acetyl-CoA Acetyl-CoA A Acetoacetyl coA Pha Lipids acetoacetyl B Pha B coA Pha Isoprenoids 3-hydroxybutyrl-coA 3Pha c C hydroxybutyrl-coA PHB A.thaliana PHB CaMV+TPP+N Rubisco+Gene+T Approaches: gene pyramiding, triple construct
Molecular farming for
proteins Antibodies Subunit vaccines
Plantibodies: in plants 1. First functional Ab:
Antibodies produced
mouse IgG1 (1989)
T1 (-heavy chain) X T2( -light chain)
T3 (F1) Nascent chains targetted to ER lumen for processing and assembly of Ab 2.ScFv:most commonly used and most successful variable regions of H and L chains are responsible for antigen binding Seq. L chain variable region + Seq. H chain variable region
synthetic gene 3.Secretory Ab (sIgA):protection against dental caries caused by S. mutans :Microbial infections at mucosal sites T1 (H)X T2 (L)X T3 (J) XT4 (Sc) Topical application on human teeth & prevented colonization of S. mutans upto 4 months. Effective as produced in murine hybridoma,effective binding to antigen.
of Hirudo medicinalis. Most potent inhibitor of thrombin-high therapeutic value in the treatment of thrombosis. Advantages over heparin -Low immunogenecity -doesnt require endogenous cofactor for activity - does not react with other proteins in the blood.
Recombinant Hirudin in Brassica napus
P-O Oleosin gene Hirudin gene
Endoprotease factor Xa cleavage site Expression in transgenic plant protein targeted to oil bodies Crush seeds Spin Oil body Remove ,spin, treat with protease
protein in aq. phase
Recombinant Vaccines Edible vaccines: It can be produced for those
