Malaria is a tropical disease that has claimed many lives over the numerous centuries gone by (reference). It usually caused by the genus Plasmodium. The vector for this parasite is a female Anopheles mosquito which transmits the pathogen through bite (reference). One of the major challenges in disease vectors is insecticide resistance. Vector-borne diseases are usually a burden in developing countries because it is not easy to reduce/control disease. In many parts of Africa, Anopheles gambiae has developed resistance towards pyrethroid insecticides which is used to control the malaria vector (Kumar, Dudley, 2007). It is feared that there are high chances of reduction in efficiency of bednets which are treated using the same insecticide. Sprays containing pyrethroids will also be useless if used because it will not have any effect on the vector since it has developed resistance.
However, P450 in A. gambiae are known to be involved in the metabolism, development and detoxification [3]. They metabolize endogenous compounds like steroids and lipids and exogenous compounds like insecticides [3]. This means P450 have been implicated in the metabolic resistance to insecticides, including pyrethroids . Therefore, studies of resistance mechanisms are important to both understanding the evolution of resistance and to minimising its impact on disease control.
This report aimed to describe the detailed bioinformatics analysis of P450 gene in Anopheles gambiae. This is done with the view of evaluating the importance of this protein in tropical health, parasitology and disease vector research. Moreover, this report will also review the various researches that have been published by other researchers on the P450 protein in A. gambiae. Since bioinformatics forms an integral part of research into tropical health, parasitology and disease vectors, this report will also review the impact of the current and future use of research technologies in these fields.
In this study an unknown DNA sequence were provided, in order to reveal as much information as possible about the nature and properties of this sequence, see below a given DNA sequence.
Mystery sequence
The following methods were used to collect data for the mystery sequence:
Firstly, a bioinformatics investigation of genomic and proteomic of the mystery sequence was applied. The DNA sequence was blasted in the NCBI website using the following parameter; nucleotides blast to get similar genome data and identify the identity of the sequence and to download FASTA files for the sequences. Following that, Blastx was used for nucleotides to translated protein sequences which were applied. Several similar genome and protein sequences were revealed.
After that similar gene and protein sequences were selected. Then, an alignment search to visualize the similarity between the sequences and to detect conserved region between them was used. Using MUSCLE alignment tool in jalview software, two separate alignments were applied one for the genes sequences and other for the protein sequences. Defaults setting were used.
Next, to understand more about the properties of this protein, FASTA files containing the sequence of interest in addition to the sequences which were selected from the blast search were loaded in BLAST2Go software. Two separate searches were done, one for the genome sequences and the other for the proteins. The programme has several parameters that could be applied to collect information about the GO terms that describes the function, biological process and cellular components of the sequence. The following steps were used to retrieve as much information as we can about this sequence. First, blast search for protein sequences with the following main sitting; for Blast Db: swissprot data base was selected, and for blast program: blastp was selected, and for genome sequences, for Blast Db: nr data base and for blast program: blastx were selected, the others were kept as default. Then a GO mapping, annotation step, InterPro scan search were performed on the default setting.
Discussion
The situation regarding the lack of information of public health means that there is a huge need to understand how the mechanisms of detoxification work. One must also identify what targets there are for the new insecticides particularly in relation to A. gambiae. There are also other factors which influence the results of the experiment where there are also insects and natural factors which come into play and which are borne about by the final results. There is also a lot of analysis in this regard especially when there is comparative bio chemical analysis which brings about different results which can also be compared and contrasted. In the test which I performed here, everything is clear and concise and the factors which influenced the experiment are there.
In the diagram described above, the GO terms describe the function and biological process of the genome and protein sequences. For example, table 1 contains significant GO terms which are ranked according to their significance; beginning with NADPH, in additional, table 2 describes the biological process. Thus, these evidences are all supportive that this protein is P450.
The structure of the P450 gene is quite the same when compared to 1amo as this also demonstrates that the 3D structure of the protein is an enzyme which contains both FAD and FMN. There is also the process of catalyisation which works accordingly in this regard.
The functional annotation of novel sequence data is a key requirement for the successful generation of functional genomics in biological research. The Blast2GO suite is known to be an useful support to these approaches, especially (but not exclusively) in nonmodel species. This bioinformatics tool is ideal genomics research because of the following: it is suitable for any species but can be also customized for specific needs, it combines high throughput with interactivity and curation, and it is user-friendly and requires low bioinformatics efforts to get it running (Ashburner, Ball
Finally one has to observe the fact that there is a considerable link between insecticides and germs in the reasoning behind these experiments. All has to be taken into context when analysing the final results which however do demonstrate that these insects are actually very much resistant to insecticides and are extremely strong in this regard.
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