Abstract: A challenge for laboratories is to provide bacterial identification and antibiotic sensitivity results within a short time. Hence, advancement in the required technology is desirable to improve timing, accuracy and quality. Even with the current advances in methods used for both phenotypic and genotypic identification of bacteria the need is there to develop method(s) that enhance the outcome of bacteriology laboratories in accuracy and time. The hypothesis introduced here is based on the assumption that the chromosome of any bacteria contains unique sequences that can be used for its identification and typing. The outcome of a pilot study designed to test this hypothesis is reported in this manuscript. Methods: The complete chromosome sequences of several bacterial species were downloaded to use as search targets for unique sequences. Visual basic and SQL server (2014) were used to generate a complete set of 18-base long primers, a process started with reverse translation of randomly chosen 6 amino acids to limit the number of the generated primers. In addition, the software used to scan the downloaded chromosomes using the generated primers for similarities was designed, and the resulting hits were classified according to the number of similar chromosomal sequences, i.e., unique or otherwise. Results: All primers that had identical/similar sequences in the selected genome sequence(s) were classified according to the number of hits in the chromosomes search. Those that were identical to a single site on a single bacterial chromosome were referred to as unique. On the other hand, most generated primers sequences were identical to multiple sites on a single or multiple chromosomes. Following scanning, the generated primers were classified based on ability to differentiate between medically important bacterial and the initial results looks promising. Conclusion: A simple strategy that started by generating primers was introduced; the primers were used to screen bacterial genomes for match. Primer(s) that were uniquely identical to specific DNA sequence on a specific bacterial chromosome were selected. The identified unique sequence can be used in different molecular diagnostic techniques, possibly to identify bacteria. In addition, a single primer that can identify multiple sites in a single chromosome can be exploited for region or genome identification. Although genomes sequences draft of isolates of organism DNA enable high throughput primer design using alignment strategy, and this enhances diagnostic performance in comparison to traditional molecular assays. In this method the generated primers can be used to identify an organism before the draft sequence is completed. In addition, the generated primers can be used to build a bank for easy access of the primers that can be used to identify bacteria.
Abstract: There is strong evidence that water channel proteins
'aquaporins (AQPs)' are central components in plant-water relations
as well as a number of other physiological parameters. We had
previously reported the isolation of 24 plasma membrane intrinsic
protein (PIP) type AQPs. However, the gene numbers in rice and the
polyploid nature of bread wheat indicated a high probability of
further genes in the latter. The present work focused on identification
of further AQP isoforms in bread wheat. With the use of altered
primer design, we identified five genes homologous, designated
PIP1;5b, PIP2;9b, TaPIP2;2, TaPIP2;2a, TaPIP2;2b. Sequence
alignments indicate PIP1;5b, PIP2;9b are likely to be homeologues of
two previously reported genes while the other three are new genes
and could be homeologs of each other. The results indicate further
AQP diversity in wheat and the sequence data will enable physical
mapping of these genes to identify their genomes as well as genetic to
determine their association with any quantitative trait loci (QTLs)
associated with plant-water relation such as salinity or drought
tolerance.
Abstract: Gamboge disorder (GD) or fruit damage by the yellow sap is a major problem in mangosteen. Mangosteen plants varied in the level of GD, from very low or non GD to low, moderate and high GD. However it was difficult to differentiate between GD and non GD plants because evaluation of the disorder is strongly influenced by environment. In this study we investigated the usefulness of primer designed from bioinformatics related to cell wall strength, termed as MCWS, to predict GD. Plant materials used were 28 mangosteen plants selected based on percentage of GD categorized as high, moderate, low and very low or non GD. The result showed that the specific DNA fragments were absent in the high GD accessions. The MCWS marker suggests as a novel polymorphic marker for GD in mangosteen as well as a marker for detect variability in mangosteen as apomictic plant.
Abstract: Before performing polymerase chain reactions (PCR), a feasible primer set is required. Many primer design methods have been proposed for design a feasible primer set. However, the majority of these methods require a relatively long time to obtain an optimal solution since large quantities of template DNA need to be analyzed. Furthermore, the designed primer sets usually do not provide a specific PCR product. In recent years, evolutionary computation has been applied to PCR primer design and yielded promising results. In this paper, a particle swarm optimization (PSO) algorithm is proposed to solve primer design problems associated with providing a specific product for PCR experiments. A test set of the gene CYP1A1, associated with a heightened lung cancer risk was analyzed and the comparison of accuracy and running time with the genetic algorithm (GA) and memetic algorithm (MA) was performed. A comparison of results indicated that the proposed PSO method for primer design finds optimal or near-optimal primer sets and effective PCR products in a relatively short time.