An improved and secreted luciferase reporter for schistosomes.
نویسندگان
چکیده
Schistosomes are multicellular parasites of humans exhibiting interesting biological adaptations to their parasitic lifestyle. Concerted and in depth analyses of these adaptations and their cell and molecular biology requires further development of molecular genetic tools in schistosomes. In the current study, we demonstrate that a Gaussia luciferase reporter leads to significantly higher levels of luciferase activity in schistosomes compared to other tested luciferases. In addition, Gaussia luciferase can be secreted into culture media enabling non-invasive analysis of reporter activity. The secretion of Gaussia luciferase should allow a variety of new experimental paradigms for schistosome studies. Comparison of biolistic and electroporation transfection methods using luciferase RNA reporters and the luciferase acitivty produced indicates that electroporation of sporocysts and schistosomula is the most efficient transfection method for the four stages analyzed. These data should facilitate additional studies in schistosomes and provide a framework for further development of DNA transfection and gene expression analysis.
منابع مشابه
A novel medium-throughput biological assay system for HTLV-1 infectivity and drug discovery
Objective(s): Here, a reporter cell line containing two reporter vectors were developed, in order to monitor the Human T-Lymphotropic Virus type1(HTLV-1) infectivity and the cell viability simultaneously. Materials and Methods: The reporter cell line was constructed by stably transfected baby hamster's kidney cell line (BHK-21), with the genomes expressing two different reporters in separate pl...
متن کاملA New Reporter Gene Technology: Opportunities and Perspectives
The paper summarizes the current status of the reporter gene technology and their basics. Reporter gene technology is widely used to monitor cellular events associated with gene expression and signal transduction. Based upon the splicing of transcriptional control elements to a variety of reporter genes, it “reports” the effects of a cascade of signaling events on gene expression inside cells. ...
متن کاملSecreted dual reporter assay with Gaussia luciferase and the red fluorescent protein mCherry
The availability of a wide range of reporter proteins, which can easily be quantitated, has had a major impact on many fields of biomedical research. In some experiments with tissue culture cells, it is necessary to control for differences in transfection efficiency and in other expression parameters. This requirement has been very conveniently met with the popular dual luciferase assay. Its di...
متن کاملDual-reporter assay using two secreted luciferase genes.
Luciferase is used as a reporter enzyme to estimate gene expression in prokaryotic or eukaryotic cells because the amount of luciferase present is correlated with light intensity in the presence of excess luciferin (1). Furthermore, its sensitivity and range of linear responses are superior to those of other typical reporters (2). When the luciferase gene containing a target promoter region is ...
متن کاملتولید هورمون رشد انسانی نوترکیب توسط سلول تخمدان هامستر چینی و بررسی فعالیت زیستی آن به روش سنجش گزارشگر ژنی
Background: Cultivated mammalian cells, because of their capacity for proper protein folding, assembly and post–translational modification, have become the dominant system for production of recombinant proteins in clinical application. Therefore, the quality and efficacy of protein can be superior when expressed in mammalian cells compared to other hosts such as bacteria. Gene reporte...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- Molecular and biochemical parasitology
دوره 155 2 شماره
صفحات -
تاریخ انتشار 2007