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Using Living Surfaces As An Artistic TableauI am looking to use genetics to paint with. I will be using the tobacco plant because it is large and easy to transform with binary gene vectors via agro bacterium. I would be looking for ways to cause deliberate pattern changes in the leaf. I will be using standard plasmid prep protocols. These recombinant plasmids will be produced in e- coli (a bacterium) and then put into agro. It is my belief that the gene vectors in the e-coli are transferred to the agro bacterium by stressing the bacterium with a slight elevated temperature while being immersed in the e-coli bearing vector. Any information or links would be very helpful and I would be glad to keep you abreast of any developments. I would be certainly giving you plants derived from my research.
TimelineWork was suspended in the fall of 2002 when the plants seeded. Hopefully research will start again soon. Tobacco Keyword Link:
http://keoz5.com/tobacco/ May 5th, 2002J Michael Moore, Georgia’s expert in tobacco supplied me with bright leaf seeds and I hope to grow my first crop of plants. By the end of the summer I hope to have the makings of a small home lab to start my genetic research. July 14th,
2001
Tobacco Plants Displaying Symptoms Caused by Secondary Infection with the Tobacco Mosaic Virus http://www.rrz.uni-hamburg.de/biologie/b_online/e35/tmvsympt.htm b-online@botanik.uni-hamburg.de Peter v. Sengbusch http://www.taoherbfarm.com/herbs/resources/tobacgrow.htm How to Grow Tobacco taoherbfarm@netidea.com Vitaly wrote: nice idea 1. for such plants, i’d use tobacco as it is large and easy to transform with agro 2. as for isolation of vectors, one would use standard plasmid prep protocols found in any molec biol manual. these recombinant plasmids are produced in e- coli (a bacterium) and then put into agro please feel free to consult with me if needed vitaly
July 13th,2001http://biobase.dk/~mundy/Translab01.htmthakuga@biobase.dk Disarmed T-DNA plasmids, in which the tumor-inducing genes are deleted, can therefore be used to insert any DNA fragment to be introduced into plant cells between the two borders. New high copy plasmids for easy cloning in E. coli and carrying different selectable marker and reporter genes have also been developed. These are based on a binary system so that the T-DNA vector only carries the border repeats into which DNA can be inserted. A second plasmid, called the helper plasmid, carries the vir genes to help mobilize the T-DNA into plants cells from the T-DNA plasmid. http://www.mindfully.org/GE/HeLa-Cells-Agrobacterium.htmvitaly.citovsky@sunysb.edu Agrobacterium tumefaciens is a soil phytopathogen that elicits neoplastic growths on the host plant species. In nature, however, Agrobacterium also may encounter organisms belonging to other kingdoms such as insects and animals that feed on the infected plants. Can Agrobacterium, then, also infect animal cells? Here, we report that Agrobacterium attaches to and genetically transforms several types of human cells. In stably transformed HeLa cells, the integration event occurred at the right border of the tumor-inducing plasmid’s transferred-DNA (T-DNA), suggesting bona fide T-DNA transfer and lending support to the notion that Agrobacterium transforms human cells by a mechanism similar to that which it uses for transformation of plants cells. Collectively, our results suggest that Agrobacterium can transport its T-DNA to human cells and integrate it into their genome. http://www.ca.uky.edu/pgel/protocols/agrobacterium/agroproto/agro-tomato.htmlrddink1@pop.uky.edu Excellent lab description of Agri implantation into tomato.http://www.rycomusa.com/aspp1998/22/1002.shtmlmatthysse@unc.edu http://www.uwrf.edu/~dc01/atumelab.html Successful transfer of the T-DNA sequence to a plant cell depends on a second set of genes located elsewhere on the plasmid, called the virulence (vir) genes, and a short DNA sequence (25 bp) at each end of the T-DNA called border sequences. It is possible to remove the genes for growth regulator and opine production and leave behind only the border regions and the vir genes. In this way the plasmid is “disarmed” and can no longer cause crown gall disease. Because the genes necessary for transformation are still present the rest of the T-DNA can still be transferred. If a gene is inserted between the border region, that gene may be transferred to the plant cell genome in the way the disease causing genes were. In this way, A. tumefaciens may be used as a gene vector to introduce new genetic material into a plant genome. Important traits such as herbicide tolerance, virus and disease resistance has been introduced in this way. http://www.arts.richmond.edu/~biology/Bgoodner.www/Homepage/Agro.html#proj1bgoodner@richmond.edu My students
and I combined these strategies by first generating mutations to identify
genes and then locating those genes using enzymes that cut DNA at specific
sequences. What makes this possible is the way we generated the mutations.
Instead of inducing a change of just one or a few components of the genetic
code, we used the insertion of a large piece of novel DNA sequence as
our mutagen (Figure 2). This novel DNA sequence carries the cutting sites
for the enzymes PacI and SwaI. If the insertion of this sequence at a
particular place on the chromosome knocks out a gene, we can identify
it. For example, we can look for strains that no longer make the essential
amino acid methionine. We now know that we have hit a gene involved in
methionine biosynthesis. We then map the gene by cutting the DNA with
PacI or SwaI and comparing the pattern of pieces with that from a normal
strain (Figure 3). July 12th, 2001 http://www.agron.missouri.edu/mnl/72/67lupotto.htmldb_request@chaco.agron.missouri.edu Maize plants were grown in the field during summer seasons 1996 and 1997, hand pollinated, and immature embryos dissected at 12 DAP, explanted on N6I medium (Lupotto and Lusardi, Maydica 33:163, 1988). For transformation, immature embryos and primary embryogenic calli were co-cultivated with Agrobacterium. The procedure adopted followed the protocol described by Hiei et al. (1994). For the infection, bacteria were grown in AB medium supplemented with each strain’s proper antibiotic selection, for three days. The bacteria were collected and diluted in low pH (pH 5.2) infection medium (LSinf) at high density (O.D. 1-1.2) in the presence of 100 uM AS. Explanted embryos were immersed in LSinf, vortexed at maximal speed 20 seconds, and incubated 10 minutes. Blotted dry embryos were subsequently co-cultivated for three days on LSD1.5, 100uM AS medium, transferred on LSD1.5 plus 250 mg/l cefotaxime (or 200 mg/l timentin) for 2 days, and tested for infection with histochemical GUS assay, or cultured for embryogenic callus induction. http://www.life.uiuc.edu/farrand/lab/index.htmlstephenf@uiuc.edu My lab is interested in the biology and the molecular biology of the plant pathogen, Agrobacterium tumefaciens. This organism causes tumors, called crown galls, on susceptible plants. Tumor induction results from the transfer of a small piece of DNA, called T-DNA, from the bacterium to the plant cell during infection. The T-DNA becomes integrated into plant cell nuclear DNA and expression of genes on this segment causes the normal plant cell to differentiate into a tumor cell. Expression of additional T-DNA genes causes the plant tumor cells to produce and secrete novel small carbon compounds called opines. In turn, Agrobacterium cells can utilize opines as sole carbon and energy sources. In the bacterium, the T-DNA and the genes for opine catabolism reside on a large, extrachromosomal virulence element called the Ti plasmid. This plasmid itself is transmissible from the bacterium to recipient bacteria by a mating mechanism called conjugation. To find out more about conjugation, opine catabolism, and plant signaling click on Research Areas in the left frame. July 7th, 2001 I am beginning to focus my study of using bio-genetics as a artistic tableau towards indoor house plants. The companies that produce such plants may know more about altering the colors and pattern. If indoor plants can be altered with agrobacterium it may be possible to do research into what may cause pattern changes in the leaf. I need find a large leafed plant that can accept new genes via agrobacterium. I need to learn exactly how you isolate genes and incorporate them into agrobacterium. June 17th, 2001 http://www.colostate.edu/programs/lifesciences/TransgenicCrops/how.html#locating http://www.hgmp.mrc.ac.uk/CCP11/tutorials.txt.html#14 May 27th, 2001 This is the beginning of my use of living surfaces as a artistic tableau. The end result will use the simplest type of surface manipulation using genetics. It may well be the wing of a fruit fly. Size is not important. What is important is the ability of changing the genetics of the organism to portray images. It may be possible to engineer agrobacterium with image designed plasmids. The leaf of the wild rice Oryza sativa may be able to display these image designed plasmids if the image designed plasmids can be inserted into Oryza sativa using agrobacterium. The image designed plasmids could also be transfered via the Brown Leaf Spot (fungus - Bipolaris oryzae) showing symptoms on Oryza sativa below RICE Oryza sativa, Complete genome known. May be a possible tableau if it accepts Agrobacterium http://plantpathology.tamu.edu/Texlab/Grains/Rice/ricetop.html Linkshttp://hordeum.oscs.montana.edu/class/CHLORLEC.html Once
isolated, characterization of plastid genome structure required restriction
and developing of overlapping restriction site maps. The plastid genome
was determined to be circular through construction of complete genome
maps. Complete chloroplast genome sequences have been reported for
maize, rice, liverwort, Chlamydomonas, Euglena and several other interesting
plant species. Virtual Cellhttp://www.life.uiuc.edu/plantbio/cell/
Complete mitochondrial genome sequenceshttp://megasun.bch.umontreal.ca/ogmp/projects/other/mt_list.html
http://hordeum.oscs.montana.edu/class/agro.htm The process of infection and gene transfer Through infection and gene transfer, A. tumefaciens accomplishes two objectives. First, it obtains a protected environment in which to grow. Second, it engineers the plant cells surrounding its colony to produce unusual amino acids which the plant cell cannot catabolize, but which the bacterium can. The bacterium creates for itself a marvelous, nutrient-rich home. From the evolutionary perspective, the genes which Agrobacterium introduces have never been observed to be sexually transmitted in nature. They are, from the plant’s perspective, an evolutionary dead end. In the short term, however, they serve the needs of the bacterium. When a plant cell is wounded, components of the hypersensitive response are induced, and several bacteriocidal compounds are released. Agrobacterium senses several of these, and moves toward their source ( The infection process is composed of: chemotaxis and initial binding at the site of wounding development of a stable attachment initiation of t-DNA excision and packaging construction of transmembrane channel movement of t-DNA to plant cell movement of t-DNA to nucleus, integration and transcription The ‘transfer’ stage of this process was pretty well disgrammed in Barbara Baker’s Science paper on pathogenesis. The array of Agrobacterium genes involved in this process sounds pretty daunting initially, but becomes more straightforward over time. The primary difference between radiobacter and its infectious relatives is the presence of a large plasmid (the Ti plasmid). The Ti plasmid carries two significant gene regions, the vir region and the tDNA. The tDna is the DNA that is transferred. Most of the genes required for infection are carried within the vir region. The chromosome carries the Chv genes. In the vir region of the plasmid are the vir A, vir B, vir C, vir D, vir E, vir g and VirH operons. As an example of the internal complexity of these, vir B contains 11 characterized open reading frames. Vir A produces the protein most directly associated with chemotaxis and sensing of plant phenolic compounds (notably acetosyringone) associated with wounding. Vir H may act as a detoxifier of the plant bacteriocidal compounds produced through the wound response. http://www.plant.uoguelph.ca/research/embryo/transfrm.htm GENETIC TRANSFORMATION OF ALFALFA The following topics are available describing our research: Objectives and Transformation Experiments Currently in Progress Publications and Abstracts on Alfalfa Transformation Graduate Student ProjectsProcedure for Genetic Transformation of Alfalfa using Agrobacterium tumefaciens Culturing Agrobacterium tumefaciens - Microbiology Tips Culturing Agrobacterium tumefaciens - Streak Plates Culturing Agrobacterium tumefaciens - Liquid Culture Agrobacterium StorageTissue Culture Media used in Transformation of Alfalfa More detailed information of Alfalfa Tissue Culture Return to Bryan McKersie’s home page Agrobacterium methodThe left side of this diagram shows how Agrobacterium is used in broad-leafed (dicots) crops such as tomatoes, potatoes, cotton and canola to introduce a new gene into a plant. Agrobacterium tumefaciens is a soil microorganism that acts as a natural genetic engineer. It can insert a piece of its DNA into the chromosome of a plant cell. Monsanto researchers discovered that for some kinds of plants, Agrobacterium was the perfect method for introducing new traits into the plant. When pieces of plant tissue were added to culture with the Agrobacterium containing the new gene, the gene could be transferred into the plant cells. These cells grow into plants with the new trait. Particle GunFor some grass-like (monocots) crops, such as corn and wheat, the Agrobacterium method doesn’t work. In these crops, researchers use a particle gun demonstrated on the right side of this diagram. In this method microscopic pellets of gold or tungsten are bathed in the DNA with the new trait. The pellets are fired from the gun into the plant cells. As the particles pass through the cell, some of the DNA is left behind. The DNA from the pellet mixes with the DNA of the cell, adding the new trait. http://www.biotechbasics.com/basics.html http://www.teleport.com/~amobb/biology/genetics.html The field of Genetics is blossoming just at the same time the Web has, so there is a lot of stuff on the Web about it. A lot of the stuff is really complex and insane, they’re not all high school level, but they’re pretty cool. Genome Sciences Department - Lawrence Berkeley National Laboratory - identifies genes and determines their functions in the context of biology and human disease through computation, expression array, mapping, bio-imaging, and bio-instrumentation. http://www.genoptix.com/technologies.html Genoptix proprietary Optophoresis technology is the only technology capable of simultaneously analyzing and isolating specific cells based on their differences at the atomic level. This exquisite level of resolution fundamentally differentiates Genoptix technology from all other current methodologies. Introduction to Fruitfly Research A basic introduction to Drosophila can be found at: http://www.ceolas.org/fly/intro.html Computer-simulated genetic experiments can be performed via Virtual FlyLab at: http://www.biologylab.awlonline.com/ http://www.biologylab.awlonline.com/trial.html Can subscribe for $7.50
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