New Approach To DNA Transfection And Genetics In Schistosome Parasites

New Approach To DNA Transfection And Genetics In Schistosome Parasites

Bibliographic Details
Main Author: Shuang , Liang
Language:English
Published: Case Western Reserve University School of Graduate Studies / OhioLINK 2015
Subjects:
Molecular Biology
Parasitology
schistosome
transfection
overexpression
promoter
Online Access:http://rave.ohiolink.edu/etdc/view?acc_num=case1424201510
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spelling ndltd-OhioLink-oai-etd.ohiolink.edu-case14242015102021-08-03T06:29:18Z New Approach To DNA Transfection And Genetics In Schistosome Parasites Shuang , Liang Molecular Biology Parasitology schistosome transfection overexpression promoter Schistosome worms of the genus Schistosoma are the causative agents of schistosomiasis, a serious global disease affecting more than 240 million people worldwide. The complex life cycle and the dearth of molecular tools have made in-depth genetic modifications of the parasitic worms challenging. In my study, I describe the use of a proprietary formulation of polyethyleneimine (PEI) as an efficacious transfection tool to introduce genetic material into schistosome parasites. I show efficient expression of exogenous genes from both heterologous and schistosome endogenous promoters in transfected worms. Using schistosome myocyte enhancer factor 2 (SmMef2), a transcriptional activator critical for myogenesis and other developmental pathways, I first describe gene overexpression as an effective method to study gene functions in schistosome parasites. In addition, while well-characterized promoters are critical to dissect and investigate molecular pathways and for the molecular engineering and manipulation of biological systems, there is extreme lack of gene regulation information in schistosomes. I use PEI-mediated plasmid transfection to define and evaluate gene expression profiles from a series of endogenous and nonendogenous promoters in schistosome parasites that are potentially useful to misexpress (underexpress or overexpress) gene product levels. Taking advantages of these new data, I overexpress schistosome genes in vivo using a strong promoter and demonstrate plasmid-based misregulation of gene expression in schistosomes, producing a clear and distinct phenotype -death. I also show that differential gene expression levels regulated by different promoters could produce distinct phenotypes, affirming the value of the importance of promoter evaluation in schistosome studies. 2015-06-03 English text Case Western Reserve University School of Graduate Studies / OhioLINK http://rave.ohiolink.edu/etdc/view?acc_num=case1424201510 http://rave.ohiolink.edu/etdc/view?acc_num=case1424201510 unrestricted This thesis or dissertation is protected by copyright: some rights reserved. It is licensed for use under a Creative Commons license. Specific terms and permissions are available from this document's record in the OhioLINK ETD Center.
collection NDLTD
language English
sources NDLTD
topic Molecular Biology
Parasitology
schistosome
transfection
overexpression
promoter
spellingShingle Molecular Biology
Parasitology
schistosome
transfection
overexpression
promoter
Shuang , Liang
New Approach To DNA Transfection And Genetics In Schistosome Parasites
author Shuang , Liang
author_facet Shuang , Liang
author_sort Shuang , Liang
title New Approach To DNA Transfection And Genetics In Schistosome Parasites
title_short New Approach To DNA Transfection And Genetics In Schistosome Parasites
title_full New Approach To DNA Transfection And Genetics In Schistosome Parasites
title_fullStr New Approach To DNA Transfection And Genetics In Schistosome Parasites
title_full_unstemmed New Approach To DNA Transfection And Genetics In Schistosome Parasites
title_sort new approach to dna transfection and genetics in schistosome parasites
publisher Case Western Reserve University School of Graduate Studies / OhioLINK
publishDate 2015
url http://rave.ohiolink.edu/etdc/view?acc_num=case1424201510
work_keys_str_mv AT shuangliang newapproachtodnatransfectionandgeneticsinschistosomeparasites
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