Mechanisms of Olfaction in Insects - ResearchSpace@Auckland ...

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List of Figures Figure 4.6: Phylogram resulting from multiple sequence alignment........................ 113 Figure 4.7: EpOR34 cDNA nucleotide sequence ..................................................... 115 Figure 4.8: Schematic representation of the consensus of the predicted TM regions from seven TM prediction programs ......................................................................... 116 Figure 4.9: Transmembrane domain prediction of EpOR34 with TMHMM 2.0 algorithms .................................................................................................................. 117 Figure 4.10: Transmembrane domain prediction of EpOR34 with TMMOD algorithms. ................................................................................................................. 117 Figure 4.11: Theoretical membrane topology of EpOR34 ....................................... 118 Figure A.1: Dose response of the empty pIB-V5 His...............................................134 Figure B.1: Chromatogram showing the elution profile of His6-EpGOBP2 from HiTrap chelating HP column. ................................................................................ 136 Figure B.2: Elution profile of His6-EpGOBP2 on Q-sepharose HP column. ........... 136 Figure C.1: ClustalX multiple sequence alignment of the moth PR clade ............... 138 Figure E.1: Multiple sequence alignment in ClustalX of E. postvittana ORs with those of five other species .......................................................................................... 145 x
List of Tables List of Tables Table 1.1: Summary of the moth pheromone receptors .............................................. 17 Table 1.2: Plant volatiles detected by E. postvittana in electroantennogram experiments from (Suckling et al., 1996). ................................................................... 28 Table 2.1: Change in fluorescence at 10 -5 M of the compounds tested with EpOR1 in the Sf9 cell assay. ......................................................................................................... 48 Table 2.2: EC50 (± standard error) and Hill slope estimates of the dose response curves of five best ligands for EpOR1 ......................................................................... 51 Table 3.1: A comparison of the EC50 values ............................................................... 74 Table 4.1: Outer and nested degenerate primers used in degenerate PCR of male E. postvittana antennal cDNA. ......................................................................................... 87 Table 4.2: Primer pairs for qRT-PCR amplification ................................................... 93 Table 4.3: Gene specific primers used for amplifying 3‟RACE products. ................. 96 Table 4.4: Primers used for amplifying 5‟ RACE products ........................................ 98 Table 4.5: Analysis of the 454 transcriptome sequence data .................................... 102 Table 4.6: E. postvittana OR repertoire to date ........................................................ 104 Table D.1: Reagents for making 1 L of Nuclei Extraction Buffer, pH 6. ................. 139 xi
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Page 3 and 4: Abstract Abstract Olfaction or the
Page 5 and 6: Acknowledgements Acknowledgements I
Page 7 and 8: Contents v Contents Abstract……
Page 9 and 10: Contents 4.2.5 Degenerate PCR .....
Page 11: List of Figures List of Figures Fig
Page 15 and 16: List of Abbreviations gDNA Genomic
Page 17 and 18: 1.1 General overview 1 General Intr
Page 19 and 20: General Introduction 3 moth in loca
Page 21 and 22: General Introduction 5 Figure 1.1:
Page 23 and 24: General Introduction 7 1.5 Componen
Page 25 and 26: General Introduction 9 OBPs have al
Page 27 and 28: General Introduction 11 A model has
Page 31 and 32: General Introduction 15 Tanaka et a
Page 33 and 34: General Introduction 17 Table 1.1:
Page 37 and 38: General Introduction 21 et al., 200
Page 39 and 40: General Introduction 23 the attract
Page 41 and 42: General Introduction 25 leaves and
Page 45 and 46: General Introduction 29 Expression
Page 47 and 48: General Introduction 31 BmOr2 HvOr2
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Page 51 and 52: Functional characterisation of Epip
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Functional characterisation of Epip
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3.1 Introduction 3 The roles of Epi
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The roles of Epiphyas postvittana G
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4.1 Introduction 4 Identification o
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Identification of putative odorant
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Microarray OR 454 Transcriptome OR
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5.1 Introduction 5 Concluding Discu
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Concluding Discussion 127 VOBA sett
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Concluding Discussion 129 redundant
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Concluding Discussion 131 against t
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Concluding Discussion 133 sequencin
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Appendix B 135 Buffered TB (Sambroo
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Appendix C 137 C. Appendix C - Clus
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Appendix D 139 D. Appendix D - Solu
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Appendix E 141 I II
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Appendix E 143 V
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Appendix E 145 VII continued Figure
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References 147 Ansebo, L., Ignell,
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References 149 Butenandt, A., Beckm
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References 151 Fedurco, M., Romieu,
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References 153 Große-Wilde, E., St
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References 155 Hrdy, I., F. Kuldova
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References 157 Kaissling, K. E. (19
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References 159 Krieger, J., Große-
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References 161 Lopez-Gomez, R. and
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References 163 Miura, N., Nakagawa,
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References 165 Pell, J. K., Macaula
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References 167 Røstelien, T., Stra
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References 169 Stowers, L. and Loga
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References 171 Turner, C. T., Davy,
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References 173 Vogt, R. G., Riddifo
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References 175 Widmayer, P., Heifet
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