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You searched for subject:(060108 Protein Trafficking). Showing records 1 – 3 of 3 total matches.

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Lincoln University

1. Xu, Janet Boyu. Cell biology and biochemical studies of ovine batten disease.

Degree: 2018, Lincoln University

The neuronal ceroid lipofuscinoses (NCLs, Batten disease) are a group of fatal inherited neurodegenerative lysosomal storage diseases of humans, which result in severe cortical atrophy, blindness, seizures, and the accumulation of fluorescent lysosome-derived organelles in neurons and most other cells throughout the body. Two naturally occurring of forms of NCL in sheep, CLN5 and CLN6, are used to study human disorders. A mutation in a soluble lysosomal protein (CLN5) causes NCL in Borderdale sheep and the South Hampshire sheep have a defective intracellular endoplasmic reticulum membrane protein (CLN6). The functions of these proteins are still unclear. Generation of highly specific and sensitive antibodies to these proteins would be very useful for biological and functional studies. Studies described in this thesis used ovine CLN5 and CLN6 NCL models to establish the full length coding sequences of the genes and expression of the encoded proteins. Antibodies against ovine CLN5 and CLN6 were generated and used for characterisation studies of both proteins in vitro. The full length ovine CLN5 gene sequence was amplified using a two-step PCR and ligation strategy with internal overhanging primers. Ovine CLN5 exon 1 has an allelic variant, but it was not associated with the CLN5 sheep mutation. Recombinant ovine CLN5 and CLN6 proteins were not expressed in Escherichia coli (E. coli) prokaryotic systems, perhaps because the gene sequences may be toxic to the host. Recombinant ovine CLN5 and CLN6 proteins were expressed successfully in eukaryotic mammalian cells using lentiviral vectors. The adenoviral antibodies against ovine CLN5 and CLN6 specifically recognized ovine CLN5 and CLN6 as shown by immunocytochemistry, Western blotting and mass spectrometry. The rabbit anti-CLN5 antibodies specifically identified the ovine mature CLN5 glycosylated form with a molecular weight of 60 kDa in media and cells and a molecular weight of 35 kDa when deglycosylated by PNGase F. The rabbit anti-CLN6 antibodies specifically detected overexpressed ovine CLN6 at molecular weight of 27 kDa in cells by Western blotting. Mass spectrometry revealed that the CLN5 N-signal sequence is cleaved off when it is mature. The deglycosylation study confirmed that CLN5 is a soluble glycosylated protein, containing high-mannose or complex type glycans at six out of eight predicted N-glycosylation sites. A lysosomal localisation of ovine CLN5 study was confirmed by rabbit anti-CLN5 antibodies labelling of the cells that also expressed the lysosomal associated membrane protein LAMP1, while ER intracellular localisation of ovine CLN6 was revealed by co-staining with rabbit anti-CLN6 antibodies and an ER marker. The CLN5 antibodies also detected endogenous CLN5 expression throughout the normal sheep and human brains. This study also provides evidence for the hypothesis of interactions between CLN5 and CLN6 in cultured and virally transduced neural cell lines using the anti-CLN5 and anti-CLN6 antibodies. Co-expression of CLN6 and CLN5 up-regulated…

Subjects/Keywords: Batten disease; neuronal ceroid lipofuscinosis; lysosomal storage disease; animal models; sheep; brain; CLN5; CLN6; proteins; antibodies; 060108 Protein Trafficking; 070709 Veterinary Pathology; 110903 Central Nervous System

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6th Edition):

Xu, J. B. (2018). Cell biology and biochemical studies of ovine batten disease. (Thesis). Lincoln University. Retrieved from http://hdl.handle.net/10182/10185

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16th Edition):

Xu, Janet Boyu. “Cell biology and biochemical studies of ovine batten disease.” 2018. Thesis, Lincoln University. Accessed December 03, 2020. http://hdl.handle.net/10182/10185.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7th Edition):

Xu, Janet Boyu. “Cell biology and biochemical studies of ovine batten disease.” 2018. Web. 03 Dec 2020.

Vancouver:

Xu JB. Cell biology and biochemical studies of ovine batten disease. [Internet] [Thesis]. Lincoln University; 2018. [cited 2020 Dec 03]. Available from: http://hdl.handle.net/10182/10185.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Xu JB. Cell biology and biochemical studies of ovine batten disease. [Thesis]. Lincoln University; 2018. Available from: http://hdl.handle.net/10182/10185

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation


University of Queensland

2. Qi, Xiaying. The role of sorting nexins in macropinocytosis and Salmonella invasion.

Degree: Institute for Molecular Bioscience, 2015, University of Queensland

Subjects/Keywords: Macropinocytosis; Sorting nexin (SNX); S. Typhimurium; Phosphoinositides; 060108 Protein Trafficking; 060109 Proteomics and Intermolecular Interactions (excl. Medical Proteomics)

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6th Edition):

Qi, X. (2015). The role of sorting nexins in macropinocytosis and Salmonella invasion. (Thesis). University of Queensland. Retrieved from http://espace.library.uq.edu.au/view/UQ:371851

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16th Edition):

Qi, Xiaying. “The role of sorting nexins in macropinocytosis and Salmonella invasion.” 2015. Thesis, University of Queensland. Accessed December 03, 2020. http://espace.library.uq.edu.au/view/UQ:371851.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7th Edition):

Qi, Xiaying. “The role of sorting nexins in macropinocytosis and Salmonella invasion.” 2015. Web. 03 Dec 2020.

Vancouver:

Qi X. The role of sorting nexins in macropinocytosis and Salmonella invasion. [Internet] [Thesis]. University of Queensland; 2015. [cited 2020 Dec 03]. Available from: http://espace.library.uq.edu.au/view/UQ:371851.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Qi X. The role of sorting nexins in macropinocytosis and Salmonella invasion. [Thesis]. University of Queensland; 2015. Available from: http://espace.library.uq.edu.au/view/UQ:371851

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

3. Shanks, Melissa Christine. Site-directed mutagenesis of the membrane-proximal (MPER) α-helical E2 region of DENV E to characterise the significance of individual residues in protein expression and migration through the secretory pathway.

Degree: School of Chemistry and Molecular Biosciences, 2015, University of Queensland

Subjects/Keywords: DENV-2; Dengue fever; Flavivirus; Virus Fusion; Structural Proteins; Envelope Proteins; 030406 Proteins and Peptides; 0601 Biochemistry and Cell Biology; 060108 Protein Trafficking

Protein Expression 2.7.1. Cell lysate preparation 2.7.2. SDS polyacrylamide gel electrophoresis… …Detection of E protein production with SDS-PAGE and Western immunoblot 5.4. Results: Detection of… …in-cell and surface expressed prME protein production with confocal microscopy 5.1.1… …Analysis of cellular and surface expressed prME protein production in transfected cells 5.1.2… …cellular and surface expressed MPER E2 mutant protein 9 CHAPTER 6 RESULTS: INSECT CELL LINE… 

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6th Edition):

Shanks, M. C. (2015). Site-directed mutagenesis of the membrane-proximal (MPER) α-helical E2 region of DENV E to characterise the significance of individual residues in protein expression and migration through the secretory pathway. (Thesis). University of Queensland. Retrieved from http://espace.library.uq.edu.au/view/UQ:363952

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16th Edition):

Shanks, Melissa Christine. “Site-directed mutagenesis of the membrane-proximal (MPER) α-helical E2 region of DENV E to characterise the significance of individual residues in protein expression and migration through the secretory pathway.” 2015. Thesis, University of Queensland. Accessed December 03, 2020. http://espace.library.uq.edu.au/view/UQ:363952.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7th Edition):

Shanks, Melissa Christine. “Site-directed mutagenesis of the membrane-proximal (MPER) α-helical E2 region of DENV E to characterise the significance of individual residues in protein expression and migration through the secretory pathway.” 2015. Web. 03 Dec 2020.

Vancouver:

Shanks MC. Site-directed mutagenesis of the membrane-proximal (MPER) α-helical E2 region of DENV E to characterise the significance of individual residues in protein expression and migration through the secretory pathway. [Internet] [Thesis]. University of Queensland; 2015. [cited 2020 Dec 03]. Available from: http://espace.library.uq.edu.au/view/UQ:363952.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Shanks MC. Site-directed mutagenesis of the membrane-proximal (MPER) α-helical E2 region of DENV E to characterise the significance of individual residues in protein expression and migration through the secretory pathway. [Thesis]. University of Queensland; 2015. Available from: http://espace.library.uq.edu.au/view/UQ:363952

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

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