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Yeasts, small eukaryotes that can be manipulated in the lab, like prokaryotes, have been studied extensively. Baker s yeast, Saccharomyces cerevisiae, has a naturally occurring plasmid. In addition, bacterial plasmids have been introduced into yeast. Unfortunately, the cells tend to lose these plasmids. This tendency has been overcome, however, by constructing bacterial plasmids that contain a yeast centromere (CEN) and the origin of yeast DNA replication (ARS for autonomously replicating sequence; g. 13.16). The yeast then carries the plasmids from one gen-
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eration to the next. The plasmids can have telomeric sequences inserted, and they can then be made linear by cutting the telomeric sequences with endonuclease. Alternatively, the plasmids can be linearized rst, and then have telomeric sequences added to their ends.The plasmids are then called yeast arti cial chromosomes (YACs). The particular advantage YACs have is that they are capable of accepting very large pieces of inserted DNA. Remember that a cosmid can hold about 50 kb; a YAC can hold as much as 800 kb or more. The ability to clone this much DNA is valuable when working with large eukaryotic genes and in the Human Genome Project (see the section with this title later in the chapter). Recombinant DNA studies in yeast have increased our knowledge about gene regulation in eukaryotes, about how the centromere works, and about the way in which the tips of eukaryotic linear chromosomes are replicated. In addition, YACs have allowed us to analyze and sequence very large segments of eukaryotic DNA.
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Tamarin: Principles of Genetics, Seventh Edition
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III. Molecular Genetics
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13. Genomics, Biotechnology, and Recombinant DNA
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synthesizing ability is lethal because the cell cannot replicate its DNA. The diaminopimelate is a cell-wall constituent; without it, the cells burst. These bacteria also carry mutations that make them extremely sensitive to bile salts. Thus, if by accident the cells were to escape, they would pose virtually no threat. The plasmids used for recombinant research were modi ed so that they could not be transferred from one cell to the next. Again, if containment failed, neither the host cells nor their plasmids would survive. In 1979, the guidelines were relaxed. Although it was wise to be cautious, it appears that initial fears were unwarranted. Recombinant DNA work now seems to pose little danger: Containment works very well, and engineered bacteria do very poorly under natural conditions. E. coli has been living in mammalian guts for millions of years, so it has had numerous opportunities to incorporate mammalian DNA into its genome (intestinal cells are dying and sloughing off into the gut all the time). No Andromeda strain has arisen, nor do we foresee one in the future.
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Current concern is focused on the acceptability of genetically modi ed crops (GM crops). As we will discuss later, one fourth of American cropland is planted with genetically modi ed crops, modi ed mainly for insect resistance. These modi cations have curtailed our use of insecticides. (For cotton and corn, for example, liquid insecticide use dropped by 3.6 million liters and powdered insecticide by 300,000 kilograms in 1999.) However, people are concerned with the effects these modi cations might have on natural ecosystems: How many valuable insects will be killed by mistake Although Third World countries are desperate for these technologies, the United States, European and Asian trading partners are demanding that the crops we export be genetically unmodi ed. Farmers are also concerned that genetically modi ed crops have been modi ed to be sterile (so called terminator technology ) so that farmers would need to buy new seeds each year. More recently, the recombinant DNA dispute has taken a whole new twist. It now has surfaced as a con ict between academic freedom and
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industrial secrecy. It seems that recombinant DNA technology is very lucrative. Numerous academic scientists have either begun genetic engineering companies or become af liated with pharmaceutical companies. However, the philosophies of private enterprise and academia are often in con ict. Academic endeavors are presumably open, with free exchange of information among colleagues, whereas private enterprise entails some degree of secrecy, at least until patents are obtained to protect the investments of the companies.Thus, a basic con ict can arise for scientists trained in gene cloning. The con ict has been prevalent since late 1980, when the rst patent for recombinant DNA techniques was awarded to Stanford University and the University of California. When, in April 2000, United States President Bill Clinton and British Prime Minister Tony Blair issued a joint statement asking that human genome data not be patented, the American stock market took a major downturn. This is a tumultuous time for biotechnology.
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