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Detecting Protein-Protein Interactions with RNA Helicase A (RHA-1) in Caenorhabditis elegans

Permanent Link: http://ncf.sobek.ufl.edu/NCFE003776/00001

Material Information

Title: Detecting Protein-Protein Interactions with RNA Helicase A (RHA-1) in Caenorhabditis elegans A Yeast Two-Hybrid Screen Assay
Physical Description: Book
Language: English
Creator: Gardner, Brent
Publisher: New College of Florida
Place of Publication: Sarasota, Fla.
Creation Date: 2007
Publication Date: 2007

Subjects

Subjects / Keywords: C. elegans
RNA Helicase A
Two-Hybrid
Genre: bibliography   ( marcgt )
theses   ( marcgt )
government publication (state, provincial, terriorial, dependent)   ( marcgt )
born-digital   ( sobekcm )
Electronic Thesis or Dissertation

Notes

Abstract: RNA Helicase A (RHA-1) is a highly conserved protein in many organisms, with orthologs in nematodes, fruit flies, mice, cows, and humans. In C. elegans, a homozygous deletion of the rha-1 gene results in stunted gonadal development, and RHA-1 is known to be involved in a variety of cellular processes from transcription regulation to nuclear transport. RHA-1 is also known to interact with BRCA-1 and Creb Binding Protein; yet, despite knockout and protein-protein interaction studies, not many other biochemical mechanisms are known concerning RHA-1 in C. elegans. The purpose of this study was to find other protein-protein interactions with RHA-1 in C. elegans using yeast two-hybrid screening methods. Once protein-protein interactions are found, more in-depth mechanisms and biological processes surrounding RHA-1 in C. elegans, and in other organisms including humans, may be inferred. The yeast two-hybrid method derives its selective screening power from the genetic separation of a transcription activator (GAL4) into its two main domains: the DNA-binding domain (DBD) and the transcription activation domain (TAD). The DBD is genetically attached to RHA-1, the bait, and the TAD is then genetically tagged to random C. elegans library proteins, the preys. When a resulting protein-protein interaction occurs, the DBD and TAD are again brought back together to constitute a complete GAL4 transcription activator once more. Resulting transcription activation induces expression of HIS3 and lacZ reporter genes, which may be readily detected. From the twelve prey proteins pulled from the assay screen, none were found to be true positive results. Such amounts of false positives is an inherent problem with two-hybrid studies. Future two-hybrid studies are recommended in which the bait would contain only specific domains of RHA-1.
Statement of Responsibility: by Brent Gardner
Thesis: Thesis (B.A.) -- New College of Florida, 2007
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. 2007 G2
System ID: NCFE003776:00001

Permanent Link: http://ncf.sobek.ufl.edu/NCFE003776/00001

Material Information

Title: Detecting Protein-Protein Interactions with RNA Helicase A (RHA-1) in Caenorhabditis elegans A Yeast Two-Hybrid Screen Assay
Physical Description: Book
Language: English
Creator: Gardner, Brent
Publisher: New College of Florida
Place of Publication: Sarasota, Fla.
Creation Date: 2007
Publication Date: 2007

Subjects

Subjects / Keywords: C. elegans
RNA Helicase A
Two-Hybrid
Genre: bibliography   ( marcgt )
theses   ( marcgt )
government publication (state, provincial, terriorial, dependent)   ( marcgt )
born-digital   ( sobekcm )
Electronic Thesis or Dissertation

Notes

Abstract: RNA Helicase A (RHA-1) is a highly conserved protein in many organisms, with orthologs in nematodes, fruit flies, mice, cows, and humans. In C. elegans, a homozygous deletion of the rha-1 gene results in stunted gonadal development, and RHA-1 is known to be involved in a variety of cellular processes from transcription regulation to nuclear transport. RHA-1 is also known to interact with BRCA-1 and Creb Binding Protein; yet, despite knockout and protein-protein interaction studies, not many other biochemical mechanisms are known concerning RHA-1 in C. elegans. The purpose of this study was to find other protein-protein interactions with RHA-1 in C. elegans using yeast two-hybrid screening methods. Once protein-protein interactions are found, more in-depth mechanisms and biological processes surrounding RHA-1 in C. elegans, and in other organisms including humans, may be inferred. The yeast two-hybrid method derives its selective screening power from the genetic separation of a transcription activator (GAL4) into its two main domains: the DNA-binding domain (DBD) and the transcription activation domain (TAD). The DBD is genetically attached to RHA-1, the bait, and the TAD is then genetically tagged to random C. elegans library proteins, the preys. When a resulting protein-protein interaction occurs, the DBD and TAD are again brought back together to constitute a complete GAL4 transcription activator once more. Resulting transcription activation induces expression of HIS3 and lacZ reporter genes, which may be readily detected. From the twelve prey proteins pulled from the assay screen, none were found to be true positive results. Such amounts of false positives is an inherent problem with two-hybrid studies. Future two-hybrid studies are recommended in which the bait would contain only specific domains of RHA-1.
Statement of Responsibility: by Brent Gardner
Thesis: Thesis (B.A.) -- New College of Florida, 2007
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. 2007 G2
System ID: NCFE003776:00001

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