Phd Thesis 33 - eCommons@USASK - University of Saskatchewan

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PPAD .......................................................Protein with Peritrophin-A domain Pssm .........................................................Position specific scoring matrix qPCR........................................................Quantitative real-time PCR RACE.......................................................Rapid Amplification <strong>of</strong> cDNA Ends rCDA........................................................Recombinant chitin deacetylase REPAT.....................................................Response to pathogen RhiNODB ................................................Sinorhizobium meliloti chitin deacetylase rMcPPAD.................................................Recombinant McPPAD RNAi........................................................RNA interference Rp.............................................................Reverse primer RT-PCR....................................................Reverse transcription-PCR SAHS .......................................................Sterile artificial hemolymph solution Se..............................................................Spodoptera exigua SEM .........................................................Scanning electron microscopy Seq cov.....................................................Sequence coverage Serpin .......................................................Serine protease inhibitor Sf..............................................................Spodoptera frugiperda siRNA ......................................................Small-interfering RNA Sl ..............................................................Spodoptera litura SlAXE......................................................Streptomyces lividans chitin deacetylase SP .............................................................Serine protease SpPGDA ..................................................Streptococcus pneumoniae chitin deacetylase TBP ..........................................................Tributyl phosphine Tc .............................................................Tribolium castaneum Tn .............................................................Trichoplusia ni TR ............................................................Tracheae TrxB .........................................................Thioredoxin reductase TUB..........................................................Tubulin UDP-GlcNAc...........................................Uridine diphosphate-N-acetylglucosamine UTR..........................................................Untranslated region Xn.............................................................Xestia c-nigrum xxiii
wt..............................................................Wild type β1, 3GLU .................................................β-1, 3-glucanase 1D.............................................................One dimensional 2D ............................................................Two dimensional xxiv
Page 1 and 2: THE MOLECULAR ARCHITECTURE OF MAMES
Page 3 and 4: "Bugs are not going to inherit the
Page 5 and 6: DEDICATION This Ph.D thesis is dedi
Page 7 and 8: ACKNOWLEDGMENTS I owe my deepest gr
Page 9 and 10: 2.1.1 Mamestra configurata ........
Page 11 and 12: 4.3.3.3.1 Alkaline phosphatase ....
Page 13 and 14: 5.3.3.4 Localization...............
Page 15 and 16: 8.2.1 Preparation of dsRNA.........
Page 17 and 18: LIST OF FIGURES Figure 1.1 The alim
Page 19 and 20: Figure 5.12 Two dimensional gel ele
Page 21 and 22: LIST OF ABBREVIATIONS Ac...........
Page 23: Ld ................................
Page 27 and 28: 1.2 The Peritrophic Matrix The pres
Page 29 and 30: Now that many PM proteins have been
Page 31 and 32: 1.2.3.1 Mechanical protection and s
Page 33 and 34: 1.2.3.3 Compartmentalization of dig
Page 35 and 36: the anterior midgut and disintegrat
Page 37 and 38: of larvae reared on the leaves of r
Page 39 and 40: 2. GENERAL MATERIAL AND METHODS Thi
Page 41 and 42: Blots were prehybridized, hybridize
Page 43 and 44: samples were mixed with loading buf
Page 45 and 46: over the m/z range 50 to 1900. LC-M
Page 47 and 48: acetylglucosamine (UDP-GlcNAc) as t
Page 49 and 50: For SEM analyses, PMs were dissecte
Page 51 and 52: Chapter 2) was conducted using tota
Page 53 and 54: significant difference was detected
Page 55 and 56: The PMs from all three stages studi
Page 57 and 58: amino acids (Figure 3.4) including
Page 59 and 60: 1 McCHS-B MATKPKTPGFTGLGDDSEDESEYTP
Page 61 and 62: 36 1 70 AiCHI MKAILATLAVLAVVTTAIEAD
Page 63 and 64: 38 1 70 AiNAG MWLQKYSLCAVYITLLSVICV
Page 65 and 66: 3.3.4 Expression Analysis of the Ge
Page 67 and 68: Figure 3.10 Expression of Mamestra
Page 69 and 70: H. virescens (Ryerse et al., 1992)
Page 71 and 72: 3.4.3 Tissue Specific Expression of
Page 73 and 74: (Chamankhah et al., 2003), McSerpin
Page 75 and 76:
4. SURVEY AND PRELIMINARY CHARACTER
Page 77 and 78:
Table 4.1 RT-PCR primers used in ex
Page 79 and 80:
Table 4.2 Proteins identified from
Page 81 and 82:
Figure 4.2 Two dimensional gel elec
Page 83 and 84:
Figure 4.3 Expression of Mamestra c
Page 85 and 86:
Trypsins Chymotrypsins Elastase 57
Page 87 and 88:
(Figure 4.5 continued) 351 420 McAP
Page 89 and 90:
(Figure 4.5 continued) 1051 1102 Mc
Page 91 and 92:
4.3.3.2.3 Insect intestinal lipases
Page 93 and 94:
4.3.3.2.5 α-Amylase Mascot analysi
Page 95 and 96:
4.3.4.3 REPAT Mascot analysis ident
Page 97 and 98:
Appendix A). All proteins were pred
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eported to be restricted to midgut
Page 101 and 102:
and Ellar, 2007; Angelucci et al.,
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the M. configurata PM. McALP1 is pr
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larvae; whereas HMG176 is expressed
Page 107 and 108:
4.4.4 Proteins without Orthologs Th
Page 109 and 110:
5.2 Material and Methods 5.2.1 Rapi
Page 111 and 112:
Table 5.2 (continued) Primer 1 Sequ
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uffer (without β-mercaptoethanol)
Page 115 and 116:
5.3.2 Insect Intestinal Mucins (McI
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proline (9.3%), histidine (11.1%),
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molting larvae (Figure 5.4B). A low
Page 121 and 122:
Figure 5.6 One dimensional (1D) and
Page 123 and 124:
(Figure 5.7 continued) 421 429 BmCD
Page 125 and 126:
meliloti (RhiNODB) and Colletotrich
Page 127 and 128:
5.3.3.5 Demonstration of chitin dea
Page 129 and 130:
similar colour changes, indicating
Page 131 and 132:
5.3.4.3 Gene expression Expression
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transfer to a gelatin impregnated p
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produced and stored in secretory ve
Page 137 and 138:
The results of CDA activity assays
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catalytically active. Of the 10 lip
Page 141 and 142:
6. SURVEY, CHARACTERIZATION AND EVO
Page 143 and 144:
PAD, conserved aromatic amino acids
Page 145 and 146:
6.2.5 Antisera Production, Protein
Page 147 and 148:
McPPAD1 MIAKFLTTVL LLNVVLTAEI PQKNA
Page 149 and 150:
with the 951 bp cDNA. The McPPAD2 p
Page 151 and 152:
The anti-rMcPPAD1 antiserum reacted
Page 153 and 154:
Four types of PPADs were identified
Page 155 and 156:
Figure 6.7 (legend is on the next p
Page 157 and 158:
Figure 6.8 Model showing the propos
Page 159 and 160:
Figure 6.9 Modeling of Mamestra con
Page 161 and 162:
6.4.1.2 Expression Genes encoding P
Page 163 and 164:
Table 6.4 (continued) Species Prote
Page 165 and 166:
however, this is not present in McP
Page 167 and 168:
that bonds could form between C2 an
Page 169 and 170:
to form a bond with C3 in the eight
Page 171 and 172:
Mamestra configurata nucleopolyhedr
Page 173 and 174:
solution. In parallel, PMs without
Page 175 and 176:
Figure 7.1 Analysis of strongly ass
Page 177 and 178:
Figure 7.3 Analysis of Mamestra con
Page 179 and 180:
kDa representing the degradation pr
Page 181 and 182:
8. TARGETING GENES INVOLVED IN PERI
Page 183 and 184:
Table 8.1 RT-PCR primers used for a
Page 185 and 186:
50 µl 0.1% Triton-100 in nuclease
Page 187 and 188:
Figure 8.1 RT-PCR analysis to detec
Page 189 and 190:
clear as the same dosages and dsRNA
Page 191 and 192:
RNAi to be initiated by dsRNA feedi
Page 193 and 194:
2003), which could inhibit the acti
Page 195 and 196:
in vitro (Cho et al., 2000), which
Page 197 and 198:
glycan, two GlcNAc residues are lin
Page 199 and 200:
The current study is also the first
Page 201 and 202:
TcCHS2 are specialized for synthesi
Page 203 and 204:
Blair, D.E., Hekmat, O., Schuttelko
Page 205 and 206:
Chapman, R.F., 1985. Structure of t
Page 207 and 208:
Dong, D-J., He, H-J., Chai, L-Q., W
Page 209 and 210:
Filho, B.P.D., Lemos, F.J.A., Secun
Page 211 and 212:
Grossi de Sa, M.F., Chrispeels, M.J
Page 213 and 214:
Huang, X., Madan, A., 1999. CAP3: A
Page 215 and 216:
Kim, M.G., Shin, S.W., Bae, K.S., K
Page 217 and 218:
Lepore, L.S., Roelvink, P.R., Grana
Page 219 and 220:
novel, very large fluorescent lipoc
Page 221 and 222:
Palli, S.R., Locke, M., 1987. Hemol
Page 223 and 224:
Ponnuvel, K.M., Nakazawa, H., Furuk
Page 225 and 226:
Schorderet, S., Pearson, R.D., Vuoc
Page 227 and 228:
Strobl, S., Maskos, K., Betz, M., W
Page 229 and 230:
Tomoyasu, Y., Miller, S.C., Tomita,
Page 231 and 232:
Wang, S., Jayaram, A.S., Hemphala,
Page 233 and 234:
Zhou, H-X., Tan, X-M., Li, C-Y., Wa
Page 235 and 236:
Appendix A. (continued) Protein Acc
Page 237 and 238:
Page 239 and 240:
Page 241 and 242:
Page 243 and 244:
218 Appendix B. Amino acid composit
Page 245 and 246:
220 Appendix D. Data from serine pr
Page 247 and 248:
Appendix F. Domain organization and
Page 249 and 250:
Appendix F. (continued) Spodoptera
Page 251 and 252:
Appendix F. (continued) The colour
Page 253 and 254:
Appendix G. (continued) 27. McPM1 C
Page 255 and 256:
Appendix G. (continued) 81. HaIIM4
Page 257 and 258:
Appendix G. (continued) 135. SeCBP6
Page 259 and 260:
234 Appendix I. Tandem repeats with
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Phd Thesis 33 - eCommons@USASK - University of Saskatchewan

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