Material Information |
| Title: |
Purification and Characterization of C. elegans Malate Dehydrogenase with the Use of Deae Ion-Exchange, Affinity and Gel Exclusion Chromatography |
| Physical Description: |
Book |
| Language: |
English |
| Creator: |
Shanks, Christopher |
| Publisher: |
New College of Florida |
| Place of Publication: |
Sarasota, Fla. |
| Creation Date: |
2008 |
| Publication Date: |
2008 |
Subjects |
| Subjects / Keywords: |
C. elegans Purification
Enzyme
Malate Dehydrogenase |
| Genre: |
bibliography ( marcgt )
theses ( marcgt )
government publication (state, provincial, terriorial, dependent) ( marcgt )
born-digital ( sobekcm )
Electronic Thesis or Dissertation |
Notes |
| Abstract: |
This thesis continued the work of previous thesis students Ira Do (2000), Danny Gonzalez (2001), and Sarah Bondi (2002). For this thesis, malate dehydrogenase of Caenorhabditis elegans (MDH-1) was expressed in Escherichia coli using the pBAD/His A vector (Invitrogen). MDH-1 was purified using a newly developed DEAE ion-exchange resin followed by a Ni-affinity column. Based on results of gel filtration chromatography, the quaternary structure of native MDH-1 appears to be a dimer. Storing MDH-1 with the coenzyme NADH did not contribute to thermostability of the enzyme as shown by kinetic assays of the enzyme. The final purified MDH-1 had fewer contaminating proteins than samples prepared in previous research projects, but was significantly less concentrated. There were also indications that MDH-1 has a higher thermostability when the enzyme is more highly purified, as shown by kinetic assays of MDH-1 samples stored in thermal denaturation conditions. However, MDH-1 appeared to lose activity after repeated, rapid changes in temperature. |
| Statement of Responsibility: |
by Christopher Shanks |
| Thesis: |
Thesis (B.A.) -- New College of Florida, 2008 |
| Electronic Access: |
RESTRICTED TO NCF STUDENTS, STAFF, FACULTY, AND ON-CAMPUS USE |
| Bibliography: |
Includes bibliographical references. |
| Source of Description: |
This bibliographic record is available under the Creative Commons CC0 public domain dedication. The New College of Florida, as creator of this bibliographic record, has waived all rights to it worldwide under copyright law, including all related and neighboring rights, to the extent allowed by law. |
| Local: |
Faculty Sponsor: Walstrom, Katherine |
Record Information |
| Source Institution: |
New College of Florida |
| Holding Location: |
New College of Florida |
| Rights Management: |
Applicable rights reserved. |
| Classification: |
local - S.T. 2008 S5 |
| System ID: |
NCFE004019:00001 |
|
Material Information |
| Title: |
Purification and Characterization of C. elegans Malate Dehydrogenase with the Use of Deae Ion-Exchange, Affinity and Gel Exclusion Chromatography |
| Physical Description: |
Book |
| Language: |
English |
| Creator: |
Shanks, Christopher |
| Publisher: |
New College of Florida |
| Place of Publication: |
Sarasota, Fla. |
| Creation Date: |
2008 |
| Publication Date: |
2008 |
Subjects |
| Subjects / Keywords: |
C. elegans Purification
Enzyme
Malate Dehydrogenase |
| Genre: |
bibliography ( marcgt )
theses ( marcgt )
government publication (state, provincial, terriorial, dependent) ( marcgt )
born-digital ( sobekcm )
Electronic Thesis or Dissertation |
Notes |
| Abstract: |
This thesis continued the work of previous thesis students Ira Do (2000), Danny Gonzalez (2001), and Sarah Bondi (2002). For this thesis, malate dehydrogenase of Caenorhabditis elegans (MDH-1) was expressed in Escherichia coli using the pBAD/His A vector (Invitrogen). MDH-1 was purified using a newly developed DEAE ion-exchange resin followed by a Ni-affinity column. Based on results of gel filtration chromatography, the quaternary structure of native MDH-1 appears to be a dimer. Storing MDH-1 with the coenzyme NADH did not contribute to thermostability of the enzyme as shown by kinetic assays of the enzyme. The final purified MDH-1 had fewer contaminating proteins than samples prepared in previous research projects, but was significantly less concentrated. There were also indications that MDH-1 has a higher thermostability when the enzyme is more highly purified, as shown by kinetic assays of MDH-1 samples stored in thermal denaturation conditions. However, MDH-1 appeared to lose activity after repeated, rapid changes in temperature. |
| Statement of Responsibility: |
by Christopher Shanks |
| Thesis: |
Thesis (B.A.) -- New College of Florida, 2008 |
| Electronic Access: |
RESTRICTED TO NCF STUDENTS, STAFF, FACULTY, AND ON-CAMPUS USE |
| Bibliography: |
Includes bibliographical references. |
| Source of Description: |
This bibliographic record is available under the Creative Commons CC0 public domain dedication. The New College of Florida, as creator of this bibliographic record, has waived all rights to it worldwide under copyright law, including all related and neighboring rights, to the extent allowed by law. |
| Local: |
Faculty Sponsor: Walstrom, Katherine |
Record Information |
| Source Institution: |
New College of Florida |
| Holding Location: |
New College of Florida |
| Rights Management: |
Applicable rights reserved. |
| Classification: |
local - S.T. 2008 S5 |
| System ID: |
NCFE004019:00001 |
|