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Section 6 : Gene Transfer

Authors: Judith A. Scheppler1, Patricia E. Cassin2, Rosa M. Gambier3
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Affiliations: 1: Grainger Center for Imagination and Inquiry, Illinois Mathematics and Science Academy, Aurora, Illinois; 2: Biology Department, Nassau Community College, Garden City, New York; 3: Biotechnology Learning Center, Suffolk County Community College, Selden, New York
Content Type: Textbook
Publication Year: 2000

Category: Applied and Industrial Microbiology

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Abstract:

Yeasts are single-cell eukaryotic organisms that contain a relatively small genome yet display many cellular and molecular features in common with higher eukaryotes. Yeasts are easy to propagate and their genome is easy to manipulate, qualities that make them a good model organism for studying eukaryotic processes and for producing eukaryotic proteins. This section first discusses the importance of using yeasts in biotechnology, explains transformation in yeasts, and compares and contrasts the uses of yeasts and bacteria. Then, it moves on to plant transformation methodologies. Plants can be genetically transformed by biological and physical methods, and transgenic plants are increasingly important in agriculture. Many dicotyledonous plants (dicots) are attacked by , a pathogen causing crown gall tumors. Tumor production is induced by the presence of the Ti (tumor-inducing) plasmid. The Ti plasmid can be genetically manipulated to serve as a vector for genes of interest. The section also describes the mechanism and role of Ti plasmid infection in plants, and the significance of opines. In addition, it lists some applications of gene transfer in plants.

Citation: Scheppler J, Cassin P, Gambier R. 2000. Gene Transfer, p 187-201. In Biotechnology Explorations. ASM Press, Washington, DC. doi: 10.1128/9781555818135.ch6
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Gene Transfer
Citation: Scheppler J, Cassin P, Gambier R. 2000. Gene Transfer, p 187-201. In Biotechnology Explorations. ASM Press, Washington, DC. doi: 10.1128/9781555818135.ch6
/content/10.1128/9781555818135.chap6.fig20-1
Figure 20.1

Synthesis of beta-carotene. Beta-carotene is synthesized from farnesyl pyrophosphate and isopentenyl pyrophosphate. This pathway requires four enzymes: geranylgeranyl pyrophosphate synthase, phytoene synthase, phytoene dehydrogenase-4H, and lycopene cyclase. When beta-carotene is synthesized, yeast colonies will be pigmented yellow. In the absence of lycopene cyclase, lycopene, a red pigment, accumulates instead.

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Figure 20.1

Synthesis of beta-carotene. Beta-carotene is synthesized from farnesyl pyrophosphate and isopentenyl pyrophosphate. This pathway requires four enzymes: geranylgeranyl pyrophosphate synthase, phytoene synthase, phytoene dehydrogenase-4H, and lycopene cyclase. When beta-carotene is synthesized, yeast colonies will be pigmented yellow. In the absence of lycopene cyclase, lycopene, a red pigment, accumulates instead.

Citation: Scheppler J, Cassin P, Gambier R. 2000. Gene Transfer, p 187-201. In Biotechnology Explorations. ASM Press, Washington, DC. doi: 10.1128/9781555818135.ch6
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Figure 20.2

Plasmids pARC145G and pARC1520, used to transform for production of beta-carotene. pARC145G contains the genes for GGPP synthase and phytoene synthase. pARC1520 contains the genes for phytoene dehydrogenase-4H and lycopene cyclase. All four gene products must be present for synthesis of beta-carotene.

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Figure 20.2

Plasmids pARC145G and pARC1520, used to transform for production of beta-carotene. pARC145G contains the genes for GGPP synthase and phytoene synthase. pARC1520 contains the genes for phytoene dehydrogenase-4H and lycopene cyclase. All four gene products must be present for synthesis of beta-carotene.

Citation: Scheppler J, Cassin P, Gambier R. 2000. Gene Transfer, p 187-201. In Biotechnology Explorations. ASM Press, Washington, DC. doi: 10.1128/9781555818135.ch6
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Citation: Scheppler J, Cassin P, Gambier R. 2000. Gene Transfer, p 187-201. In Biotechnology Explorations. ASM Press, Washington, DC. doi: 10.1128/9781555818135.ch6
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Figure 21.1

The Ti plasmid. T-DNA includes left and right borders and auxin, cytokinin, and opine synthesis genes. Other sequences on the plasmid include the ori (origin of replication), genes, and opine catabolism genes.

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Image of Figure 21.1
Figure 21.1

The Ti plasmid. T-DNA includes left and right borders and auxin, cytokinin, and opine synthesis genes. Other sequences on the plasmid include the ori (origin of replication), genes, and opine catabolism genes.

Citation: Scheppler J, Cassin P, Gambier R. 2000. Gene Transfer, p 187-201. In Biotechnology Explorations. ASM Press, Washington, DC. doi: 10.1128/9781555818135.ch6
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Figure 21.2

Chemical structure of nopaline, an opine.

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Figure 21.2

Chemical structure of nopaline, an opine.

Citation: Scheppler J, Cassin P, Gambier R. 2000. Gene Transfer, p 187-201. In Biotechnology Explorations. ASM Press, Washington, DC. doi: 10.1128/9781555818135.ch6
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/content/10.1128/9781555818135.chap6.fig21-3
Figure 21.3

Guide diagram for paper chromatography of leaf lysates.

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Image of Figure 21.3
Figure 21.3

Guide diagram for paper chromatography of leaf lysates.

Citation: Scheppler J, Cassin P, Gambier R. 2000. Gene Transfer, p 187-201. In Biotechnology Explorations. ASM Press, Washington, DC. doi: 10.1128/9781555818135.ch6
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Figure 21.4

Paper chromatography apparatus. (A) Paper chromatogram cylinder showing positions of samples and staples. (B) After equilibrium of liquid and gas phases, the chromatogram is inserted in the closed chamber. (C) A glass plate is used to cover and seal the chamber.

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Image of Figure 21.4
Figure 21.4

Paper chromatography apparatus. (A) Paper chromatogram cylinder showing positions of samples and staples. (B) After equilibrium of liquid and gas phases, the chromatogram is inserted in the closed chamber. (C) A glass plate is used to cover and seal the chamber.

Citation: Scheppler J, Cassin P, Gambier R. 2000. Gene Transfer, p 187-201. In Biotechnology Explorations. ASM Press, Washington, DC. doi: 10.1128/9781555818135.ch6
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References

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