wellcome trust research career development fellowship: a personal perspective samantha sampson...
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Wellcome Trust Research Career Development Fellowship:
A personal perspective
Samantha SampsonDepartment of Microbiology
Centre for Molecular Microbiology and Infection
Career History
Cape Town, South Africa:• BSc (Biochemistry & Chemistry)• BSc Hons, MSc & PhD (Medical Biochemistry)• Mycobacterium tuberculosis strain diversity
WT RCDF• Opportunity for postdoctoral scientists to become independent research scientists and to undertake research of high quality• Eligibility:
• 3-6 years research experience from date of PhD degree• intellectual contributions to published research• demonstrate potential to carry out independent research
WT RCDF• Application Procedure:
• Preliminary application form (25/9/08)• Full application (to appropriate funding committee)• Panel Interview
TB: Disease Burden
• 1/3rd of world’s population infected with Mycobacterium tuberculosis• 8 million incident cases• 2 million deaths annually
PE/PPE proteins
PPE-MPTR subgroup (up to ~3720 aa)
(GGA/N GGA/N)n
PE-PGRS subgroup (up to >1500 aa)
PE
NXGXGNXG
PPE
PPE-SVP subgroup(up to ~470 aa)
GXXSVPXXWPPEPE
“PE-only” subgroup (~110 aa)
Subgroup with unique C-terminal region (up to ~560 aa)
Subgroup with unique C-terminal region (up to ~600 aa)
PE PPE
The proposal
• Title: The PE and PPE proteins of Mycobacterium tuberculosis and Mycobacterium bovis: exploring their potential as variable antigens
• Hypothesis: PE and PPE genes of M. tuberculosis and M. bovis encode variable antigens
The proposal
Aims: (i) Establish whether the PE and PPE genes are differentially expressed under different in vitro growth conditions and during host infection.
(ii) Determine whether PE and PPE epitopes are differentially recognised by the host immune system over the course of an infection.
(iii) Elucidate mechanisms of PE and PPE gene regulation.
Kinetics of PE/PPE immune responses>PE35MEKMSHDPIAADIGTQVSDNALHGVTAGSTALTSVTGLVPAGADEVSAQAATAFTSEGIQLLASNASA...MEKMSHDPIAADIGTQVSDN IAADIGTQVSDNALHGVTAG VSDNALHGVTAGSTALTSVT VTAGSTALTSVTGLVPAGAD TSVTGLVPAGADEVSAQAAT AGADEVSAQAATAFTSEGIQ QAATAFTSEGIQLLASNASA...
37 ºC, 5% CO2, 48 hours
Store plasma @ -80 ºC until use
PE/PPE regulation
Labeled probe (0.8 ng Rv1195 upstream region)
Unlabeled probe
Protein (100 ng, rRv0485-His6)
+
-
-
1
+
-
+
2
+
+
3
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4
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5
DNA-proteincomplex
Unbound, labelled DNA
Lung organ burden
0 25 50 75 100 125 150 175103
104
105
106
107
108
H37Rv
0485
Time (days)
Lo
g10
CF
U
Balb/cJ mouse survival
0 100 200 300 400 5000
102030405060708090
100 M. tuberculosis H37Rv
M. tuberculosis H37Rv:0485
Time (days)
Per
cen
t su
rviv
al
Why was my application successful?
• WT Remit: “… research across all biomedical science … to produce knowledge that can be used to protect and improve human and animal health.”• Preliminary data• Sponsor• Institution• Track record• Preparation• Right place at the right time!
Independent funding: Pros
• Independence• Limited teaching obligations (if you choose)• Technical assistance• Opportunity to develop own research interests• Develop management skills (time, budget, personnel)
Advice
• Be flexible (personally and professionally)• Be pro-active• Have a Plan B• Research your options• DO WHAT YOU ENJOY!!
Acknowledgments
• Prof. Rob Warren (PhD Supervisor)
• Prof. Barry Bloom (Post-doc PI)
• Prof. Douglas Young (RCDF Sponsor)
• Rachael Goldstone (RA)