dc.contributor.author |
Arnett, Diana. |
en_US |
dc.contributor.author |
Youngstown State University. Dept. of Biology. |
en_US |
dc.date.accessioned |
2011-01-31T14:17:29Z |
|
dc.date.accessioned |
2019-09-08T02:27:48Z |
|
dc.date.available |
2011-01-31T14:17:29Z |
|
dc.date.available |
2019-09-08T02:27:48Z |
|
dc.date.created |
2000 |
en_US |
dc.date.issued |
2000 |
en_US |
dc.identifier |
45268492 |
en_US |
dc.identifier.other |
b18647856 |
en_US |
dc.identifier.uri |
http://jupiter.ysu.edu/record=b1864785 |
en_US |
dc.identifier.uri |
http://hdl.handle.net/1989/6151 |
|
dc.description |
1 v. (various pagings). : ill. ; 29 cm. |
en_US |
dc.description |
Thesis (M.S.)--Youngstown State University, 2000. |
en_US |
dc.description |
Includes bibliographical references (leaves 84-87). |
en_US |
dc.description.abstract |
The quinic acid (qa) gene cluster is a positively regulated system. In the
absence of the inducer, quinic acid, the repressor protein binds to the activator
protein, blocking transcription of the qa genes. Addition of quinic acid releases the
activator protein, which is then free to bind to its activator binding sites, increasing
the levels of transcription of the qa genes. The activator binding sites are composed
of a 16 base pair (bp) conserved sequence, but the importance of the individual bases
in the sites is currently unknown.
To determine the importance of the bases, the DNA containing the activator
binding site was cloned and isolated. The -127 binding site of the qa-2 gene was
chosen due to its high binding affinity for the activator protein. A plasmid known to
contain the entire qa cluster was digested with Pst! and the fragment containing the
qa-2-qa-x intergenic region was cloned into pBR322 to form plasmid 177. This was
digested further with the enzymes EcoRI and Pst! and the desired' fragment cloned
into pBluescript to form pEP, which was then digested with EcoRI and HindUI and
the appropriate fragment was cloned into pBluescript to form pEH. PEH was finally
digested with KpnI to give a 500 bp fragment that was cloned into M13. Using the
method described by Kunkel, et al (1991), site-directed mutagenesis was performed
on one of the most highly conserved bases in the site. A clone was then sequenced to
determine if the mutagenesis was successful. While the clone chosen did not
contained the desired mutation, future students will study the remaining prepared to
clones to attempt to isolate a mutant. |
en_US |
dc.description.statementofresponsibility |
by Diana Arnett. |
en_US |
dc.language.iso |
en_US |
en_US |
dc.relation.ispartofseries |
Master's Theses no. 0700 |
en_US |
dc.subject.classification |
Master's Theses no. 0700 |
en_US |
dc.subject.lcsh |
Neurospora crassa. |
en_US |
dc.title |
Site-directed mutagenesis of the -127 activator binding site of the qa-2 gene of neurospora crassa / |
en_US |
dc.type |
Thesis |
en_US |