BIOL125_Ch10 Text Notes

BIOL125_Ch10 Text Notes - BIOL 125 (Microbiology) Text...

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BIOL 125 (Microbiology) Text Notes Chapter 10: Genetic Engineering 10.1. BASIC ELEMENTS AND APPLICATIONS OF GENETIC ENGINEERING a. Genetic Engineering – also called bioengineering , is the direct, deliberate modification of an organism’s genome. 10.2. TOOLS AND TECHNIQUES OF GENETIC ENGINEERING a. Practical Properties of DNA i. Enzymes for Dicing, Splicing, and Reversing Nucleic Acids 1) Restriction Endonucleases – can clip polynucleotide strands of DNA crosswise at selected positions. a) These enzymes originate in bacterial cells; recognize foreign DNA; and are capable of breaking the phosphodiester bonds between adjacent nucleotides on both strands of DNA. b) Lecture Notes: Names of the enzymes are given based on the bacteria from which it was isolated. 2) Palindromes – sequences of DNA that are identical when read from 5’ to 3’ direction to one strand and the 3’ to 5’ direction on the other strand. 3) Endonucleases are used in the lab to cut DNA into smaller pieces for further study as well as to remove and insert it during recombinant DNA techniques. 4) Restriction Fragments – pieces of DNA produced by restriction endonucleases. a) Restriction Fragment Length Polymorphisms (RFLP) restriction fragments of different lengths. 5) Ligase – enzyme used in the final splicing of genes into plasmids and chromosomes. It rejoins the phosphate-sugar bonds cut by endonucleases. a) Lecture Notes: Used for generating recombinant DNA molecules. 6) Reverse Transcriptase – used for converting RNA to DNA. 7) Complementary DNA (cDNA) – copies made from mRNA, tRNA, rRNA, and other forms of RNA. ii. Analysis of DNA 1) Gel Electophoresis – one way to produce a readable pattern of DNA fragments. a) Lecture Notes: Gel electrophoresis separates DNA fragments based on size. File: 5afff90bbd0dfe5be12f127d6fcbfc52b88896a5.doc Updated 8/14/09
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Microbiology Text Notes Chapter 10 Page 2 i) DNA samples are placed on soft agarose gel and subjected to an electric current. Negatively charged DNA then move toward the positive pole. ii) Rate of movement is dependent on the size of the fragment. The larger fragments move slower. iii) Fragments are stained using EtBr for observation iii. Nucleic Acid Hybridization and Probes 1) Two different nucleic acids can hybridize by uniting at their complementary site. a) Hybridization – process that matches complementary strands of nucleic acid (DNA-DNA, RNA-DNA, RNA-RNA). Used for locating specific sites or identifying exact sequences of nucleic acids (Text Glossary). 2) Gene Probes – oligonucleotide tracers which consist of a short stretch of DNA of a known sequence that will base-pair with a stretch of DNA with a complementary sequence, if one exists in the test sample. a)
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BIOL125_Ch10 Text Notes - BIOL 125 (Microbiology) Text...

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