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--- chapter_12 [2025/04/11 11:11] – [Using an insertion library to find interesting mutants] mike
+++ chapter_12 [2025/04/15 06:57] (current) – [Creating an insertion library for yeast] mike
@@ Line 47: / Line 47: @@
   * A library of yeast genomic fragments cloned into a bacterial plasmid. We learned about the concept of genomic libraries in [[chapter_09|Chap. 09]].
   * The //E. coli// $lacZ$ gene. We learned about the $lacZ$ gene in Chapters [[chapter_09|09]] and [[chapter_10|10]]. In this experiment the $lacZ$ gene is going to be used in yeast cells as a reporter gene (sometimes just called a reporter) for transcriptional activity of yeast genes. The $lacZ$ coding sequence works in yeast because //E. coli// and yeast both use the exact same universal genetic code for converting triplet codon sequences into amino acids.
-  * A modified bacterial transposon called mini-Tn7 (Fig. {{ref>Fig3}}). Transposons are naturally occurring pieces of DNA that can transpose, or jump around, to random locations in genomes. We can modify transposons such that we can experimentally control when they jump around, and we can also construct them to carry genetic markers that help us track the transposon. In this experiment, we have engineered mini-Tn7 to contain the $lacZ$ gene (but without any cis-acting regulatory sequences such as $lacO$ or $lacP$), a yeast gene called $URA3$ required for uracil prototrophy (in this case we are including native upstream regulatory sequences necessary for yeast cells to express $URA3$, and an //E. coli// gene (plus appropriate bacterial regulatory sequences) that confers drug resistance to the antibiotic tetracycline ($tet^R$).
+  * A modified bacterial transposon called mini-Tn7 (Fig. {{ref>Fig3}}). Transposons are naturally occurring pieces of DNA that can transpose, or jump around, to random locations in genomes. We can modify transposons such that we can experimentally control when they jump around, and we can also construct them to carry genetic markers that help us track the transposon. In this experiment, we have engineered mini-Tn7 to contain the $lacZ$ gene (but without any cis-acting regulatory sequences such as $lacO$ or $lacP$), a yeast gene called $URA3$ required for uracil prototrophy (in this case we are including native upstream regulatory sequences necessary for yeast cells to express $URA3$), and an //E. coli// gene (plus appropriate bacterial regulatory sequences) that confers drug resistance to the antibiotic tetracycline ($tet^R$).
 <figure Fig3>