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The expression and activity of the Type IV pili in Streptococcus
Taiwan (FMS-Taiwan) - Chang Gung University, Chinese Dpt, Taipei
Department of Microbiology & Immunology
Type of Research Project
- Basic science
What is the background of the project?
Streptococcus sanguinis is a primary colonizer of the dental biofilm and an opportunistic pathogen for subacute endocarditis. Interestingly, this species is the only known streptococcal species that harbors an operon for type IV pili (Tfp) biosynthesis. Our earlier studies have found that Tfp participate in the adherence and biofilm formation of S. sanguinis, and in some strains, Tfp also drive surface-dependent motility. Although the organization of the operon is very similar among strains, the number of the pilA gene in the pil cluster, encoding the structural subunit (pilin) of Tfp, varies from strains to strains. Thus, this project is designed to investigate whether the differences in the pilA genes affect the functions of Tfp, and how the differences may affect the virulence of Streptococcus sanguinis.
What is the aim of the project?
To understand the role of the type IV pili in the pathogenesis of Streptococcus sanguinis.
What techniques and methods are used?
1. DNA sequence analysis: various sequence analysis tools including Vector NTI, BLAST, domain search analysis will be used. 2. Western analysis: to verify pilin protein production 3.Microscopy: both transmission electron microscopy and confocal laser scanning microscope (CLSM) will be used to examine Tfp and biofilm
What is the role of the student?
- The student will observe the practical experiments but will be highly involved in the analysis of the results
- The tasks will be done under supervision
What are the tasks expected to be accomplished by the student?
The student will first examine the twitching motility of 80 clinical isolates. Based on the motility phenotype, isolates with different levels of motility will be selected. The sequence of the pilA gene from the selected isolates will be obtained by PCR followed by sequence analysis. The basic characteristics of the obtained sequences will be analysed by various sequence analysis tools. The Tfp phenotypes of these isolates will be observed by TEM with immune-gold labelling with anti-PilA antibody. Biofilm formation of the selected isolates in a flow cell system will be analysed and examined by CLSM. The student is required to attend weekly laboratory meeting and present progress. At the end of the internship, the student will give an oral presentation and a written report is also required.
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
The PI will explain the project and other information to the study. One technician will guide the student one the bench.
What is expected from the student at the end of the research exchange? What will be the general outcome of the student?
- The student will prepare a presentation - The student will prepare a scientific report - The student’s name will be mentioned in a future publication - The student will have the opportunity to present the results together with the supervisor at a conference
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
Basic knowledge of Basic bacteriology.
Are there any legal limitations in the student’s involvement
Type of students accepted
This project accepts: - Medical students - Graduated students (less than 6 months) - Dental medicine students (IADS members)
- Appl Environ Microbiol. 2019 Mar 6;85(6). pii: e02788-18. doi: 10.1128/AEM.02788-18.
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