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WEB TUTORIAL 15.1 Producing the Bovine Growth Hormone Text Sections Section 15.2 Transgenic Biotechnology, p. 234 Introduction How do scientists use recombinant DNA technology to produce a protein such as bovine growth hormone, which can then be used to increase milk production in cows? In this tutorial, you’ll see how bacteria can be used as factories for the production of human or other animal proteins. Learning Objectives • • Understand what a recombinant plasmid is. Know the basic steps involved in producing recombinant proteins in bacteria. Narration Cloning Genes Using Bacteria Each cell of a cow has all of the DNA needed to make all of the cow's proteins, including bovine growth hormone, or BGH. This DNA is found in the chromosomes in each cell's nucleus. To clone the BGH gene, DNA is taken from the cow cell's nucleus and cut with a restriction enzyme that leaves “sticky ends” on either side of the BGH gene. “Sticky ends” are so-named because they are able to base pair with any DNA molecule containing a complementary sticky end. The BGH gene will be cloned by putting it into a bacterial plasmid. A plasmid is a circular piece of bacterial DNA that normally exists separate from the bacterial chromosome and can replicate itself. In order to insert the BGH gene into the plasmid, the plasmid must be removed from the bacterial cell and cut open with the same restriction enzyme used on the cow DNA. When the pieces of cow DNA and the cut plasmid DNA are mixed together, their complementary sticky ends will join, forming recombinant plasmids. After the recombinant plasmid has been formed it is put back into a bacterial cell, which forms many copies, or clones, of the recombinant plasmid as it divides. Once inside the bacteria, the BGH gene can be expressed, i.e., the bacterial cell uses the information contained in the inserted BGH gene to produce BGH protein (called recombinant BGH or rBGH). The rBGH is easily separated from the bacteria and recovered from the media in which the bacteria are grown. You should now be able to… • • • Compare the experiments that Redi and Pasteur used to address the possibility of spontaneous generation and specify the experimental variables being tested in their experiments. Explain the importance of a control in an experiment. Describe the effects of boiling on microbial growth.