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Purification and Characterization of C. elegans Malate Dehydrogenase with the Use of Deae Ion-Exchange, Affinity and Gel...

Permanent Link: http://ncf.sobek.ufl.edu/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

Permanent Link: http://ncf.sobek.ufl.edu/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

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