Haemolymph clotting in Drosophila melanogaster and Galleria mellonella [Elektronische Ressource] / submitted by Christoph Scherfer

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Informations

• Publié par: justus-liebig-universitat_giessen
• Publié le: 01 janvier 2004
• Nombre de lectures: 34
• Langue: English
• Poids de l'ouvrage: 53 Mo

Extrait

Haemolymph clotting in
Drosophila melanogaster
and Galleria mellonella
A Thesis submitted for the Degree of Doctor of Natural Sciences
(Dr. rer. nat.) at the Faculty of Biology, Chemistry and Geosciences (FB08),
Justus-Liebig-University Gießen, Germany
Inaugural-Dissertation zur Erlangung des Doktorgrades der
Naturwissenschaften (Dr. rer nat.)
des Fachbereiches Biologie, Chemie und Geowissenschaften (FB08)
der Justus-Liebig-Universität Gießen, Deutschland
submitted by / vorgelegt von
Christoph Scherfer
Gießen 2004______________________________________________________________________
The experimental work presented in this study has been conducted at the Institute for
Molecular Biology and Functional Genomics at Stockholm University (Sweden) from
January 2001 to August 2004. It was performed under the supervision of Prof. Dr. Tina
Trenczek (Institute for General and Special Zoology, Justus-Liebig-University Gießen,
Germany) and Docent Dr. Ulrich Theopold (Institute for Molecular Biology and
Functional Genomics, Stockholm University, Sweden). The work was supported by a
PhD fellowship (No. D/01/41725) of the German Academic Exchange Service (DAAD)
from May 2001 to April 2004.
This study will be electronically published and publicly available for download at
http://geb.uni-giessen.de/geb/.
Die praktischen Arbeiten der vorliegenden Studie wurden am Institut für
Molekularbiologie und Funktionsgenomik an der Universität Stockholm (Schweden)
von Januar 2001 bis August 2004 durchgeführt. Sie wurde von Prof. Dr. Tina Trenczek
(Institut für Allgemeine und Spezielle Zoologie, Fachbereich 08 der Justus-Liebig-
Universität Gießen, Deutschland) und Dozent Dr. Ulrich Theopold (Institut für
Molekularbiologie und Funktionsgenomik, Universität Stockholm, Schweden) betreut.
Die Arbeit wurde von Mai 2001 bis April 2004 durch ein Promotionsstipendium
(Kennziffer: D/01/41725) des Deutschen Akademischen Austauschdienstes (DAAD)
unterstützt.
Diese Studie wird elektronisch veröffentlicht werden und unter der Adresse
http://geb.uni-giessen.de/geb/ zum Download zur Verfügung stehen.
Dekan: Prof. Dr. Jürgen Mayer
1. Gutachterin: Prof. Dr. Tina Trenczek
2. Gutachter: Dozent Dr. Ulrich Theopold______________________________________________________________________
Statement
With this I state that the present work, to the best of my knowledge and belief, does not
contain material previously published or written by another person, except where due
reference has been made in the text.
Eidesstattliche Erklärung
Hiermit erkläre ich, die vorliegende Arbeit selbständig durchgeführt und verfasst und
keine anderen als die angegebenen Hilfsmittel verwendet zu haben.
___________________________
Gießen, September 2004______________________________________________________________________
The Blind Men and the Elephant
(John Godfrey Saxe, 1816-1887, based on an Indian fable)
It was six men of Indostan The Fifth, who chanced to touch the ear,
To learning much inclined, Said: “E`en the blindest man
Who went to see the Elephant Can tell what this resembles most;
(Though all of them were blind), Deny the fact who can
That each by observation This marvel of an Elephant
Might satisfy his mind Is very like a fan!”
The First approached the Elephant The Sixth no sooner had begun
And happening to fall About the beast to grope,
Against his broad and sturdy side, Then, seizing on the swinging tail
At once began to bawl: That fell within his scope,
“God bless me! but the Elephant “I see,” quoth he, “the Elephant
Is very like a wall!” Is very like a rope!”
The Second, feeling of the tusk, And so these men of Indostan
Cried, “Ho! what have we here Disputed loud and long,
So very round and smooth and sharp? Each in his own opinion
To me `tis mighty clear Exceeding stiff and strong,
This wonder of an Elephant Though each was partly right,
Is very like a spear!” And all were in the wrong!
The Third approached the animal,
And happening to take
The squirming trunk within his hands,
Thus boldly up and spake:
“I see,” quoth he, “the Elephant
Is very like a snake!” Moral:
The Fourth reached out an eager hand, So oft in these scientific wars,
And felt about the knee. The disputants, I ween,
“What most this wondrous beast is like Rail on in utter ignorance
Is mighty plain,” quoth he; Of what each other mean,
`Tis clear enough the Elephant And prate about an Elephant
Is very like a tree!” Not one of them has seen!______________________________________________________________________
Publications
Original Papers:
Li, D., Scherfer, C., Korayem, A., Zhao, Z., Schmidt, O. and Theopold, U. (2002)
Insect hemolymph clotting: evidence for interaction between the coagulation
system and the prophenoloxidase activating cascade.
Insect Biochem. Molec. Biol. 32, 919-928.
Scherfer, C., Karlsson, C., Loseva, O., Bidla, G., Goto, A., Havemann, J.,
Dushay, M. S. and Theopold, U. (2004)
Isolation and characterization of hemolymph clotting factors in Drosophila
melanogaster by a novel pull-out assay. Current Biology 14, 625-629.
Karlsson, C., Korayem, A. M., Scherfer, C., Loseva, O., Dushay, M. S. and
Theopold, U. (2004)
Proteomic analysis of the Drosophila larval hemolymph clot.
Submitted to Journal of Biological Chemistry.
Korayem, A. M., Fabbri, M., Takahashi, K., Scherfer, C., Lindgren, M., Schmidt, O.,
Ueda, R., Dushay, M. S. and Theopold, U. (2004)
A Drosophila salivary gland mucin is also expressed in immune tissues: evidence
for a function in coagulation and the entrapment of bacteria.
Submitted to Insect Biochemistry and Molecular Biology.
Review article:
Theopold, U., Li, D., Fabbri, M., Scherfer, C., Schmidt, O. (2002)
The coagulation of insect hemolymph. Cell. Mol. Life Sci. 59, 363-372.______________________________________________________________________
Abbreviations
ALP alkaline phosphatase
Bc Black cells
BCIP 5-bromo-4-chloro-3-indolyl phosphate
bp base pairs
cDNA complementary deoxyribonucleic acid
DAB 3',3'-diaminobenzidine tetrachloride
dig UTP digoxygenin-modified UTP
DMSO dimethylsulfoxide
DNA deoxyribonucleic acid
dom domino (D. melanogaster)
dsRNA double-stranded RNA
DTT dithiothreitol
ECM extracellular matrix
EDTA ethylenediamine tetraacetic acid
EGTA ethyleneglycol-bis (beta-aminoethylether)-
N,N,N`,N`-tetraacetic acid
EST expressed sequence tag
FACS fluorescence-activated cell sorter
fbp1 fat body protein 1 (D. melanogaster)
FCS fetal calf serum
FITC fluorescein isothiocyanate
g gram
g gravity [in italics]
Gal4 yeast transcription factor involved in expression of
galactose-induced genes
GFP green fluorescent protein
h hour(s)
H O dd double distilled water2
hml hemolectin (D. melanogaster)
hmu hemomucin ()
HPL Helix pomatia lectin
HS hybridisation solution
IGEPAL CA-630 octylphenyl-polyethylene glycol (detergent)
IPTG isopropylthio-b-D-galactoside
kDa kilo Dalton
KGD lysine-glycine-aspartate
K half-maximal reaction rate (enzymes)m
l litre(s)
LPS lipopolysaccharide
LRE leucine-arginine-glutamate
lsp1c larval serum protein 1c (D. melanogaster)
M molar
mA milliampere______________________________________________________________________
mbn-2 tumourous cell-line derived from the haematopoietic organ of the
Drosophila mutant malignant blood neoplasm-2 (mbn-2)
MDC monodansylcadaverine
mg milligram
MHC major histocompatibility complex
min minute(s)
ml millilitre(s)
mM millimolar
mSSL mouse strictosidine synthase-like
mg microgram
ml microlitre(s)
mM micromolar
NBT Nitro blue tetrazolium chloride
NFkB nuclear factor kB
ng nanogram
Ni-NTA nickel nitrotriacetic acid
nm nanometer (wavelength)
PBS phosphate-buffered saline
PBST phosphate-buffered saline including 0,1 % Tween-20
PCR polymerase chain reaction
PGRP peptidoglycan recognition protein
PI propidium iodide
PNA peanut agglutinin (Arachis hypogaea lectin)
ppm parts per million
PTU phenylthiourea
RGD arginine-glycine-aspartate
RNA ribonucleic acid
RNAi RNA interference
RNase ribonuclease
RP49 ribosomal protein 49 (D. melanogaster)
RT-PCR reverse transcription polymerase chain reaction
SDS sodium dodecyl sulphate
sec second(s)
serpin serine protease inhibitor
SL2 Schneider’s Line S2 (Drosophila haemocyte-like cell line)
SSC sodium chloride - sodium citrate buffer
SSL strictosidine synthase-like
TBS Tris-buffered saline
TBST Tris-buffered saline including 0,1 % Tween-20
tig tiggrin (D. melanogaster)
Tris Tris-hydroxymethyl-aminomethane
tRNA transfer RNA
Tween-20 polyoxyethylenesorbitan monolaurate
UAS upstream activation sequence (from yeast promoter)
VWD von Willebrand disease
vWF von Willebrand factorTable of Contents 1
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Table of Contents
1 INTRODUCTION 3
1.1 Innate and adaptive i