900 resultados para Genotyping by sequencing


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The soda process was the first chemical pulping method and was patented in 1845. Soda pulping led to kraft pulping, which involves the combined use of sodium hydroxide and sodium sulfide. Today, kraft pulping dominates the chemical pulping industry. However, about 10% of the total chemical pulp produced in the world is made using non-wood material, such as bagasse and wheat straw. The soda process is the preferred method of chemical pulping of non-wood materials, because it is considered to be economically viable on a small scale and for bagasse is compatible with sugarcane processing. With recent developments, the soda process can be designed to produce minimal effluent discharge and the fouling of evaporators by silica precipitation. The aim of this work is to produce bagasse fibres suitable for papermaking and allied applications and to produce sulfur-free lignin for use in specialty applications. A preliminary economic analysis of the soda process for producing commodity silica, lignin and pulp for papermaking is presented.

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This work is concerned with the genetic basis of normal human pigmentation variation. Specifically, the role of polymorphisms within the solute carrier family 45 member 2 (SLC45A2 or membrane associated transporter protein; MATP) gene were investigated with respect to variation in hair, skin and eye colour ― both between and within populations. SLC45A2 is an important regulator of melanin production and mutations in the gene underly the most recently identified form of oculocutaneous albinism. There is evidence to suggest that non-synonymous polymorphisms in SLC45A2 are associated with normal pigmentation variation between populations. Therefore, the underlying hypothesis of this thesis is that polymorphisms in SLC45A2 will alter the function or regulation of the protein, thereby altering the important role it plays in melanogenesis and providing a mechanism for normal pigmentation variation. In order to investigate the role that SLC45A2 polymorphisms play in human pigmentation variation, a DNA database was established which collected pigmentation phenotypic information and blood samples of more than 700 individuals. This database was used as the foundation for two association studies outlined in this thesis, the first of which involved genotyping two previously-described non-synonymous polymorphisms, p.Glu272Lys and p.Phe374Leu, in four different population groups. For both polymorphisms, allele frequencies were significantly different between population groups and the 272Lys and 374Leu alleles were strongly associated with black hair, brown eyes and olive skin colour in Caucasians. This was the first report to show that SLC45A2 polymorphisms were associated with normal human intra-population pigmentation variation. The second association study involved genotyping several SLC45A2 promoter polymorphisms to determine if they also played a role in pigmentation variation. Firstly, the transcription start site (TSS), and hence putative proximal promoter region, was identified using 5' RNA ligase mediated rapid amplification of cDNA ends (RLM-RACE). Two alternate TSSs were identified and the putative promoter region was screened for novel polymorphisms using denaturing high performance liquid chromatography (dHPLC). A novel duplication (c.–1176_–1174dupAAT) was identified along with other previously described single nucleotide polymorphisms (c.–1721C>G and c.–1169G>A). Strong linkage disequilibrium ensured that all three polymorphisms were associated with skin colour such that the –1721G, +dup and –1169A alleles were associated with olive skin in Caucasians. No linkage disequilibrium was observed between the promoter and coding region polymorphisms, suggesting independent effects. The association analyses were complemented with functional data, showing that the –1721G, +dup and –1169A alleles significantly decreased SLC45A2 transcriptional activity. Based on in silico bioinformatic analysis that showed these alleles remove a microphthalmia-associated transcription factor (MITF) binding site, and that MITF is a known regulator of SLC45A2 (Baxter and Pavan, 2002; Du and Fisher, 2002), it was postulated that SLC45A2 promoter polymorphisms could contribute to the regulation of pigmentation by altering MITF binding affinity. Further characterisation of the SLC45A2 promoter was carried out using luciferase reporter assays to determine the transcriptional activity of different regions of the promoter. Five constructs were designed of increasing length and their promoter activity evaluated. Constitutive promoter activity was observed within the first ~200 bp and promoter activity increased as the construct size increased. The functional impact of the –1721G, +dup and –1169A alleles, which removed a MITF consensus binding site, were assessed using electrophoretic mobility shift assays (EMSA) and expression analysis of genotyped melanoblast and melanocyte cell lines. EMSA results confirmed that the promoter polymorphisms affected DNA-protein binding. Interestingly, however, the protein/s involved were not MITF, or at least MITF was not the protein directly binding to the DNA. In an effort to more thoroughly characterise the functional consequences of SLC45A2 promoter polymorphisms, the mRNA expression levels of SLC45A2 and MITF were determined in melanocyte/melanoblast cell lines. Based on SLC45A2’s role in processing and trafficking TYRP1 from the trans-Golgi network to stage 2 melanosmes, the mRNA expression of TYRP1 was also investigated. Expression results suggested a coordinated expression of pigmentation genes. This thesis has substantially contributed to the field of pigmentation by showing that SLC45A2 polymorphisms not only show allele frequency differences between population groups, but also contribute to normal pigmentation variation within a Caucasian population. In addition, promoter polymorphisms have been shown to have functional consequences for SLC45A2 transcription and the expression of other pigmentation genes. Combined, the data presented in this work supports the notion that SLC45A2 is an important contributor to normal pigmentation variation and should be the target of further research to elucidate its role in determining pigmentation phenotypes. Understanding SLC45A2’s function may lead to the development of therapeutic interventions for oculocutaneous albinism and other disorders of pigmentation. It may also help in our understanding of skin cancer susceptibility and evolutionary adaptation to different UV environments, and contribute to the forensic application of pigmentation phenotype prediction.

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Inclusions of sp-hybridised, trans-polyacetylene [trans-(CH)x] and poly(p-phenylene vinylene) (PPV) chains are revealed using resonant Raman scattering (RRS) investigation of amorphous hydrogenated carbon (a-C:H) films in the near IR – UV range. The RRS spectra of trans-(CH)x core Ag modes and the PPV CC-H phenylene mode are found to transform and disperse as the laser excitation energy ћωL is increased from near IR through visible to UV, whereas sp-bonded inclusions only become evident in UV. This is attributed to ћωL probing of trans-(CH)x chain inhomogeneity and the distribution of chains with varying conjugation length; for PPV to the resonant probing of phelynene ring disorder; and for sp segments, to ћωL probing of a local band gap of end-terminated polyynes. The IR spectra analysis confirmed the presence of sp, trans-(CH)x and PPV inclusions. The obtained RRS results for a-C:H denote differentiation between the core Ag trans-(CH)x modes and the PPV phenylene mode. Furthermore, it was found that at various laser excitation energies the changes in Raman spectra features for trans-(CH)x segments included in an amorphous carbon matrix are the same as in bulk trans-polyacetylene. The latter finding can be used to facilitate identification of trans-(CH)x in the spectra of complex carbonaceous materials.

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For robots to operate in human environments they must be able to make their own maps because it is unrealistic to expect a user to enter a map into the robot’s memory; existing floorplans are often incorrect; and human environments tend to change. Traditionally robots have used sonar, infra-red or laser range finders to perform the mapping task. Digital cameras have become very cheap in recent years and they have opened up new possibilities as a sensor for robot perception. Any robot that must interact with humans can reasonably be expected to have a camera for tasks such as face recognition, so it makes sense to also use the camera for navigation. Cameras have advantages over other sensors such as colour information (not available with any other sensor), better immunity to noise (compared to sonar), and not being restricted to operating in a plane (like laser range finders). However, there are disadvantages too, with the principal one being the effect of perspective. This research investigated ways to use a single colour camera as a range sensor to guide an autonomous robot and allow it to build a map of its environment, a process referred to as Simultaneous Localization and Mapping (SLAM). An experimental system was built using a robot controlled via a wireless network connection. Using the on-board camera as the only sensor, the robot successfully explored and mapped indoor office environments. The quality of the resulting maps is comparable to those that have been reported in the literature for sonar or infra-red sensors. Although the maps are not as accurate as ones created with a laser range finder, the solution using a camera is significantly cheaper and is more appropriate for toys and early domestic robots.

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We consider boundary layer flow of a micropolar fluid driven by a porous stretching sheet. A similarity solution is defined, and numerical solutions using Runge-Kutta and quasilinearisation schemes are obtained. A perturbation analysis is also used to derive analytic solutions to first order in the perturbing parameter. The resulting closed form solutions involve relatively complex expressions, and the analysis is made more tractable by a combination of offline and online work using a computational algebra system (CAS). For this combined numerical and analytic approach, the perturbation analysis yields a number of benefits with regard to the numerical work. The existence of a closed form solution helps to discriminate between acceptable and spurious numerical solutions. Also, the expressions obtained from the perturbation work can provide an accurate description of the solution for ranges of parameters where the numerical approaches considered here prove computationally more difficult.