Abstract: . . .                                                              2005 HHMI Background Information for Laboratory 3         Plasmids were originally found in relatively simple organisms, like bacteria, algae and yeasts. Plasmids are usually much smaller than chromosomes and vary in size from a few thousand bp (a few kb) to . . .
. . .  generation vector and contains several improvements over first generation vectors such as pBR322, the map of which is on pp. 310 - 311 of the same  Biolabs catalog. Improvements in p UC19 include its smaller size (2,686 bp), a 54 bp polylinker with cloning sites for 21 restriction enzymes, and a short segment of DNA containing the regulatory sequences and the coding information for the first 146 amino acids of the ß -galactosidase ( LacZ ) gene of E. coli. The polypeptide encoded by this region . . .
. . .  plasmid with an inserted fragment actually do contain such a fragment.   2. Would a recombinant plasmid with more than one inserted fragment be more or less likely to occur than a recombinant plasmid with just one inserted fragment? Explain your answer briefly.                                                                 . . .
. . .  subunit of ß -galactosidase and is the basis for an easy assay to determine whether a foreign DNA fragment has been inserted into the polylinker .       p RAS2 , the second plasmid you will work with today, is a recombinant plasmid which was created by inserting the RAS2 gene from the genome of Baker’s yeast, Saccharomyces cerevisiae, into p UC19 so the RAS2 gene could be sequenced and studied more easily. Ras genes are of great interest in molecular biology and medicine . . .
. . .  (REs), which, as you have learned, recognize specific DNA sequences, usually 4 to 8 bp in length, and cleave at these sequences. Nathans, Smith and Arber were awarded the Nobel Prize in 1979 for discovering restriction enzymes and having the insight and creativity to use these enzymes to map genes.    . . .
. . .  carries an inserted fragment of DNA?   6. What is the purpose of using IPTG and XGal in the selective plates? Is IPTG essential for this procedure? Why or why not?   Laboratory 4: Cloning Your Plasmid and Genomic DNAs, Part III (Friday Morning)     Objectives of Laboratory 4, Part III: 1. Analyze the different types of colonies on your transformation plates 2. Count the colonies on your transformation plates . . .
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