"Pharming" and "plantibodies"
An increasingly viable option is the production of highly valuable enzymes by plants and animals. In addition to production of human proteins in these organisms (see a subsequent section), other valuable proteins that are currently produced by microorganisms could very well be produced by higher organisms instead. Animal and plant "bioreactors" in some respects may be superior to recombinant bacterial systems, because eukaryotes deliver proteins in the generally more desirable glycosylated form. These organismal systems may also be cheaper than cell fermentation techniques.
Two examples of production of human proteins in plants include the production of human serum albumin in transgenic tobacco and potato, and production of human insulin by tobacco. In both cases, the produced protein appears to be fully effective in humans. Unfortunately, however, one cannot raise his/her insulin level by eating transgenic tobacco leaves, as the protein in most cases will be broken down to amino acids before it reaches the blood stream. Therefore, in these cases one cannot escape the practice of protein isolation and purification before transgenic leaves are converted into drugs.
Over the last few years, also antibodies have been produced in plants. Initially, the antibody's light and heavy chains were produced in different plants. But a subsequent cross of these two varieties resulted in progeny carrying assembled and functional antibodies. These "plantibodies" are now used for diagnostic and therapeutic purposes (see http://www.epicyte.com/home.html and http://www.molecularfarming.com/plantigens.html for some recent examples and information).
Reversible male sterility in plants
As has been indicated earlier, heterozygous individuals often are healthier and stronger than homozygous ones. The only way to guarantee heterozygicity in plants is to make sure self-pollination cannot occur. For most crop plants it was very tedious or practically impossible to exclude selfing. To exclude self-pollination, it would be good to introduce male sterility in plants: progeny from such plants are then expected to be 100% heterozygous (assuming they were pollinated with pollen from an unrelated variety). To introduce male sterility, a promoter was identified that was turned on exclusively in tapetum cells (a tissue around the pollen sac that is essential for pollen production). This promoter then was linked up to a gene coding for a bacterial ribonuclease (named barnase). This ribonuclease selectively chops up ribonucleic acids. The promoter/ribonuclease construct was then introduced into plants (canola, tobacco, you name it). Because the promoter allows expression only in tapetum cells, the gene construct disrupts only development of the tapetal tissue and its end product, pollen. Plants transformed with this construct were male-sterile but otherwise normal.
Although male-sterile plants are valuable for hybrid seed production, they have limited value when it comes to crop production. Fertility must be restored to crops such as wheat, rice, and tomato, in which the seed or fruit is the harvested product. Fortunately, the ribonuclease is inhibited very much by a simple protein, named barstar. One can thus cross the male-sterile plant with a male-fertile variety in which the gene for barstar has been introduced, and the result is progeny with viable pollen and restored fertility.
Antisense RNA
Antisense RNA refers to nucleotide strands that are produced in a cell and that are complementary to a particular mRNA. Antisense RNA can be produced, for example, by inverting the coding region of a gene with respect to its promoter. The antisense RNA can hybridize with its corresponding mRNA, making it double-stranded. The double-stranded mRNA no longer can be recognized by the protein-synthesizing machinery (the ribosomes), and thus expression of this mRNA is suppressed. Also, in many systems double-stranded mRNA is very unstable and is broken down quickly. Thus, one can inactivate specific genes while not interfering with others. This is very useful for both agriculture and medicine, and the first examples of practical applications of this RNAi (RNA interference) technology are appearing. As with any new technology, the initial pilot projects are sort of pedantic (including manipulation of flower color and of the speed of fruit ripening). However, more exciting application possibilities abound. In the mean time, antisense technology is contributing to the birth of a new field, reverse genetics. With this method one can turn off genes, and see what the consequences are.
Antisense approaches already are used to protect plants from damage by plant viruses. For example, reversal of a gene from bean yellow mosaic virus (BYMV), and putting it into tobacco under a reasonably strong promoter, has led to a tobacco variety that is quite resistant to BYMV. A similar approach is used to transfer viral resistance to other plants. This finding is of significance, in that currently no effective, environmentally friendly methods exist to control many plant viruses.
Agricultural applications in developing countries
Perhaps indicative of the large potential and relative ease of genetic engineering, developing countries (particularly China) are progressing rapidly in development and application of genetically engineered crops. Some have gone into commercial production well ahead of similar crops in the US. In China, for example, tomatoes that have been engineered for improved virus resistance have been on the market since late 1992. There are two main reasons for the more rapid commercialization of bioengineered crops in the developing world: (1) less tight governmental approval mechanisms, and (2) hungrier populations. While in developed countries the main value of biotechnological applications may be to reduce production costs, in the developing world a main factor is the production of more food. Indeed, genetic engineering applications seem to be pretty successful to cut down on pathogen-induced losses. For example, genetic modification of papaya plants (expression of the ringspot virus coat protein in the plant) protects very well against the very destructive ringspot virus.
http://photoscience.la.asu.edu/photosyn/courses/BIO_343/
This web site contains the syllabus for the MBB 343/BIO 343 course, describes the laboratory experiments, and covers materials that will be discussed in the lecture but that have not been covered sufficiently in the required textbook (B.R. Glick and J.J. Pasternak (1998) Molecular Biotechnology: Principles and Applications of Recombinant DNA, second edition).
Most of the material at this website has been compiled over the years by Wim Vermaas, a Professor in the Plant Biology Department. Instructors this year are Wim Vermaas (weeks 5-8, 12-13, and 15-16) and David Rhoads (weeks 1-4, 9-11, and 14).
Contact information:
Wim Vermaas: office location: LS-E549; phone: (480) 965-3698; Email: wim@asu.edu; office hours for Fall 2002: M and TH 10:40-11:30.
David Rhoads: office location:LS-E511; phone (480) 965-2583; Email: david.rhoads@asu.edu; office hours for Fall 2002: T/TH 1:30-2:30 pm on the days that he lectures only.
Material at this site may be used freely by others as long as the source of the information is acknowledged. Contact the appropriate web master to learn about the rules of using materials contained at other web sites that are referred to at this site.