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Concluding Discussion 132 a wide range of temperatures. The availability of a draft genome of E. postvittana will be the first genome for a pest member of this family and will become the model organism for tortricid pests. The availability of the genome sequence of E. postvittana should enable functional genomic studies of the moth; provide a wealth of information on genes involved in biological processes and in the regulation of gene expression as well as facilitate the study of the complexity of gene families. Studies of host-plant specialisation for example, E. postvittana is polyphagous so genes involved in its survival on such a wide host range, as well as proteins involved in food digestion in the gut of the larvae can be identified. Other target processes include the regulation of genes involved in the survival of E. postvittana, such as for juvenile hormone, which have implications in physiological processes of moths can also be studied in detail. Other genes associated with the olfactory system and their regulation mechanisms could also be studied in detail. One of the major drawbacks of using chemical insecticides is development of resistance by the insects. These resistance mechanisms can also be studied and better insecticide targets developed. With the availability of genomic data for more moths, the evolution of ORs can be studied. An improved set of ORs from lepidopterans can perhaps be used in phylogenetic studies to test for associations, if any, between phylogenetic relatedness and odorant binding. The problem with any ab initio whole genome assembly is the feasibility of the assembly process. E. postvittana does not have any closely related genome sequence available for mapping its genome to. The low cost of sequencing with high throughput sequencing platforms has the drawback of producing significantly short reads that are impossible to assemble ab initio. Another problem facing E. postvittana genome assembly is that its genome size is unknown. B. mori genome has been shown to have large repeats and it is highly likely to be the case for E. postvittana. Assembling such a genome will be highly challenging. Fortunately for the genome sequencing project, a bacterial artificial chromosome (BAC) library of E. postvittana has already been constructed. BAC-end sequencing will enable local, smaller assemblies of individual BACs which can be used further to assemble longer scaffolds, thereby facilitating the assembly of the draft genome. Several BACs that potentially have genes for several of the putative ORs have already been identified. These BACs can be used as the starting point for BAC shotgun
Concluding Discussion 133 sequencing and assembly. The genome sequence dataset that we currently have has lots of gaps, as shown by the high number of scaffolds, approximately 843000 scaffolds. This coverage is being improved by further sequencing of the genome. Another complementary approach to BAC sequencing is mate pair end sequencing (Illumina, 2010), which will again give the ends of 5kb long fragments that can be used as a guide for assembling the shotgun sequencing fragments. The challenges faced within the context of this thesis in identifying new ORs from E. postvittana is reflected by the low levels of homology between the members of this group and the importance of high throughput sequencing technologies to such projects. With the identification and decoding of increasing number of moth ORs, important questions about correlation of the phylogenetic relatedness of these proteins with their ligands, as well as the occurrence and significance of multiple PRs in moths can be addressed.
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Abstract Abstract Olfaction or the
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Acknowledgements Acknowledgements I
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Contents v Contents Abstract……
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Contents 4.2.5 Degenerate PCR .....
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List of Figures List of Figures Fig
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List of Tables List of Tables Table
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List of Abbreviations gDNA Genomic
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1.1 General overview 1 General Intr
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General Introduction 3 moth in loca
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General Introduction 5 Figure 1.1:
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General Introduction 7 1.5 Componen
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General Introduction 9 OBPs have al
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General Introduction 11 A model has
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General Introduction 15 Tanaka et a
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General Introduction 17 Table 1.1:
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General Introduction 21 et al., 200
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General Introduction 23 the attract
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General Introduction 25 leaves and
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General Introduction 29 Expression
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General Introduction 31 BmOr2 HvOr2
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General Introduction 33 for lepidop
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Functional characterisation of Epip
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3.1 Introduction 3 The roles of Epi
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4.1 Introduction 4 Identification o
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Page 163 and 164: References 147 Ansebo, L., Ignell,
Page 165 and 166: References 149 Butenandt, A., Beckm
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Page 169 and 170: References 153 Große-Wilde, E., St
Page 171 and 172: References 155 Hrdy, I., F. Kuldova
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Page 181 and 182: References 165 Pell, J. K., Macaula
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Page 185 and 186: References 169 Stowers, L. and Loga
Page 187 and 188: References 171 Turner, C. T., Davy,
Page 189 and 190: References 173 Vogt, R. G., Riddifo
Page 191 and 192: References 175 Widmayer, P., Heifet
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