Chapter 16 - BSC 1010 Dr. Presley Chapter 16 BIOTECHNOLOGY...

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BSC 1010 Dr. Presley Chapter 16 BIOTECHNOLOGY Biotechnology is the use of natural biological processes to: a. produce a useful product b. achieve desired result -examples: use yeast to make alcohol, bread develop turkey with lots of white meat develop disease-resistant crops 1980's -- bioengineered organisms produced using recombinant DNA technology for medical or industrial use - cloning genes, drugs, vaccines, clean up oil spills insecticides, herbicides, freeze-resistance, improved nutritional content - See Table 16.1, p. 331 and Table 16.2, p. 333 ** potential for human gene therapy RECOMBINANT DNA TECHNOLOGY Application of restriction endonucleases, Vectors Transformation Used to: locate and sequence DNA alter and assemble recombinant DNA amplify DNA fragments Goal is to produce a specific protein of interest RECOMBINANT DNA = rDNA -combine DNA from 2 different sources -uses techniques of sequencing, rejoining, amplifying, and locating DNA fragments all of which use complementary base pairing A to T (or U) and G to C - can be made in a test tube using restriction enzymes to cut the DNA - DNA ligase is the enzyme used to seal the DNA backbones together - Study Figure 16.4 on page 321 RESTRICTION ENZYMES (100+) -also called restriction endonucleases -naturally occur in some bacteria -function in bacteria is to "restrict" viral replication by cutting vDNA – thus protecting the bacterial cell from viral takeover - shown in Fig. 16.1 p. 318 -each cleaves DNA at a specific site on each DNA strand = restriction site or also called the recognition sequence -notice the palindrome nature of the restriction site (reads the same from 5’ to 3’ direction on both strands -shown in text page 318 -produce " sticky ends " which can bind to complementary sticky ends of foreign DNA cut by the same restriction enzyme - use the enzyme DNA ligase to join sugar-phosphate backbones of the recombinant DNA - examples: EcoRI, BamHI GEL ELECTROPHORESIS -method used to separate or purify DNA fragments -commonly an agarose gel is placed in a chamber with selected buffer -sample DNA fragments cut by restriction enzyme(s) is placed in a sample well -an electrical field is applied -DNA migrates to the positive end because of attraction of phosphates (neg.) -the porous gel acts as a sieve allowing smaller fragments to move faster down the field -gels are stained for visualization of banding pattern of DNA fragments in gel -type of information obtained by electrophoresis: - size of DNA fragments - localization of specific DNA sequences by a probe -pure DNA fragment can be extracted by cutting the gel - DNA can be transferred to a membrane by a “blotting” technique for further study or retrieval - This technique is used for “DNA Fingerprinting” or RFLPs (restriction fragment length polymorphism) *important tool in forensic science used in criminal cases Ch 16 1 of 7
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BSC 1010 Dr. Presley GETTING NEW GENES INTO CELLS by Vectors: VECTORS: Study Fig 16.5 on page 322 - carry foreign DNA for rDNA
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Chapter 16 - BSC 1010 Dr. Presley Chapter 16 BIOTECHNOLOGY...

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