ISSN 2736-1756
Research Article
Advanced Journal of Microbiology Research Vol. 2003
Available online at http://internationalscholarsjournals.org/journal/ajmr
© 2003 International Scholars Journals
Molecular distribution of gypsy-like retrotransposons in cotton Gossypium Spp.
Essam A. Zaki1* and Abdel Ghany A. Abdel Ghany2
1Genetic Engineering and Biotechnology Research Institute, GEBRI, Research Area, Borg El Arab, Post Code 21934, Alexandria
2Institute of Efficient Productivity, Zagazig University, Zagazig, Egypt.
Accepted 18 April, 2003
Abstract
PCR primers specific for conserved domains of the reverse transcriptase (RT) genes of gypsy-like retrotransposons amplified their corresponding gene in two Gossypium barbadense cultivars. Analysis with the FASTA software showed a high DNA sequence homology to pine, gypsy LTR-retrotransposon. Using the PCR product as a hybridization probe, gypsy-like retrotransposons were detected in wild type species of Gossypium, suggesting that gypsy-like retrotransposons are present in the Gossypium genome. This supports the view that gypsy-like retrotransposons are major components of plant genomes. Our results suggest gypsy-like retrotransposons have played a fundamental role in the shaping and evolution of the Gossypium genome.
Key words: Gossypium, gypsy, polyploidy, retroelements, retrotransposons, retroviruses, reverse transcriptase.
Essam A. Zaki*, Abdel Ghany A. Abdel Ghany
Page: 1 - 10
Research Article
Advanced Journal of Microbiology Research Vol. 2003
Available online at http://internationalscholarsjournals.org/journal/ajmr
© 2003 International Scholars Journals
Four gene introduction methods affect the shoot regeneration and localization of transgene expression in greenhouse stem explants and in vitro-grown chrysanthemum stem thin cell layers
J. A. Teixeira da Silva* and S. Fukai
Faculty of Agriculture, Kagawa University, Miki-cho, Kagawa, 761-0795, Japan
Accepted 18 April 2003
Abstract
Gene introduction method (GIM) affected shoot regeneration capacity (SRC) in standard and spray-type chrysanthemums. SRC was both cultivar and GIM-dependent in both in vitro and greenhouse stem explants, the former significantly higher than the latter. Sonication had an SRC-stimulating effect on in vitro explants. Other GIMs (Agrobacterium, biolistics, Agrolistics) had an SRC-inhibiting effect on greenhouse explants. Genotype-dependence of SRC was observed in both in vitro and greenhouse material. SRC is influenced by the explant and regeneration media, which should be modified if altered by the GIM. Shoots derived from all GIM treatments showed normal growth under in vitro and greenhouse conditions, and flowered normally. In addition, this study further shows that explant origin (in vitro versus greenhouse) and cultivar significantly affect the regeneration process, even when an optimized medium is utilized. The integration of the GUS transgene is also GIM-dependent, but in all cases is shown to occur in the venation.
Keywords: Agroinfection, biolistics, explant survival, regeneration, sonication.
J. A. Teixeira da Silva*, S. Fukai
Page: 1 - 10
Research Article
Advanced Journal of Microbiology Research Vol. 2003
Available online at http://internationalscholarsjournals.org/journal/ajmr
© 2003 International Scholars Journals
Genetic affinities of Fusarim spp. and their correlation with origin and pathogenicity
Mohmed S. Khalil1, Mohmed A. Abdel-Sattar2, Ibrahim N. Aly2, Kamel A. Abd-Elsalam1,3* and Joseph A. Verreet3
1Agricultural Research Center, Plant Pathology Research Institute, Giza, Egypt.
2Suez Canal University, Faculty of Agriculture, Ismailia, Egypt.
3Christian Albrechts Universität zu Kiel, Institut für Phytopathologie, Kiel, Germany.
Accepted 14 April, 2003
Abstract
Random amplified polymorphic DNA (RAPD) analyses was used in combination with pathogenicity assays to study the taxonomic kinships among five Fusarium species. A total of 46 isolates of Fusarium spp. obtained from diseased cotton seedlings showing typical root rot and dampping-off symptoms were characterized. Of 10 primers tested, four primers produced polymorphic amplification patterns with taxon-specific bands, in addition to individual- specific bands. Genetic analysis indicated into 2 main clusters, with the minor cluster included all F. moniliforme and F. solani at the genetic similarity of GS=57.82%. The major cluster consisted of all F. oxysporum, F. avenaceum and F. chlamydosporum clustered at 71% similarity. There was no clear-cut relationship between clustering in the RAPD dendrogram, pathogenicity test and geographic origin of tested isolates. The results suggest that RAPD-PCR is a useful method for analysing genetic variation within and between Fusarium spp.
Key words: DNA-fingerprinting, Fusarium chlamydosporum, genetic homology, RAPD-PCR.
Ibrahim N. Aly, Kamel A. Abd-Elsalam* and Joseph A. Verreet, Mohmed S. Khalil, Mohmed A. Abdel-Sattar
Page: 1 - 10
Research Article
Advanced Journal of Microbiology Research Vol. 2003
Available online at http://internationalscholarsjournals.org/journal/ajmr
© 2003 International Scholars Journals
Substrate Channelling and Energetics of Saccharomyces cerevisiae DSM 2155 Grown on Glucose in Fed-Batch Fermentation Process
Olusegun Peter AKINYEMI1, Eriola BETIKU2+*, and Bamidele Ogbe SOLOMON2
1Chemical and Polymer Engineering Department, Lagos State University, Lagos State, Nigeria.
2Chemical Engineering Department, Obafemi Awolowo University, Ile-Ife, Osun State, Nigeria.
Accepted 10 April, 2003
Data collected during high-cell-density cultivation of Saccharomyces cerevisiae DSM 2155 on glucose in a simulated five-phase feeding strategy of fed-batch process, executed on the Universal BIoprocess CONtrol (UBICON) system using 150L bioreactor over a period of 24h have been analysed. The consistency of the data set was checked using both the available electron and carbon balances. Estimates of the true energetic yields and cell maintenance requirements were obtained through the application of a multivariate statistical procedure known as covariate adjustment technique. A low value of maintenance coefficient, me = 0.004h-1, and a high average value of the true biomass energetic yield, max = 0.745, were obtained for the bioreactor system, which showed that the organism was in no danger of ethanol produced during this cultivation. A simple model for estimating the distribution of substrate consumed between the fermentative and the respiratory pathways in the oxido-reductive process was developed based on the respiratory quotient (RQ) values. The fraction of substrate consumed for respiratory metabolic activities (qsresp/qs) was virtually 1.0 for the first three phases of the feeding strategy, which accounted for the first sixteen hours of the 24h operation. This was an indication that ethanol formation was avoided during this period.
Key Words: Saccharomyces cerevisiae DSM 2155, available electron and carbon balances, fed-batch, respiratory quotient, true energetic yields, maintenance requirement.
and Bamidele Ogbe SOLOMON, Eriola BETIKU+*, Olusegun Peter AKINYEMI
Page: 1 - 10
Research Article
Advanced Journal of Microbiology Research Vol. 2003
Available online at http://internationalscholarsjournals.org/journal/ajmr
© 2003 International Scholars Journals
Bioinformatic tools and guideline for PCR primer design
Kamel A. Abd-Elsalam
Molecular Markers Lab., Plant Pathology Research Institute, Agricultural Research Center, Orman 12619, Giza, Egypt
Accepted 28 April 2003
Abstract
Bioinformatics has become an essential tool not only for basic research but also for applied research in biotechnology and biomedical sciences. Optimal primer sequence and appropriate primer concentration are essential for maximal specificity and efficiency of PCR. A poorly designed primer can result in little or no product due to non-specific amplification and/or primer-dimer formation, which can become competitive enough to suppress product formation. There are several online tools devoted to serving molecular biologist design effective PCR primers. This review intends to provide a guide to choosing the most efficient way to design a new specific-primer by applying current publicly available links and Web services. Also, the purpose here is to provide general recommendations for the design and use of PCR primers.
Key words: Bio-computing, primer design, web-based resources.
Kamel A. Abd-Elsalam
Page: 1 - 10
Research Article
Advanced Journal of Microbiology Research Vol. 2003
Available online at http://internationalscholarsjournals.org/journal/ajmr
© 2003 International Scholars Journals
A home made kit for plasmid DNA mini-preparation
Simeon Oloni KOTCHONI1 , Emma Wanjiru GACHOMO2, Eriola BETIKU3, and Olusola Olusoji SHONUKAN4
1Department of Plant Molecular Biology, Institute of Botany, Kirschallee 1, University of Bonn, D-53115 Germany.
2Institute for Plant Diseases, Nussallee 9, University of Bonn, D-53115 Germany.
3Chemical Engineering Department, Faculty of Technology, Obafemi Awolowo University, Ile-Ife, Nigeria.
4Department of Microbiology, Faculty of Science, Obafemi Awolowo University, Ile-Ife, Nigeria.
Accepted 26 March 2003
Abstract
Many methods have been used to isolate plasmid DNA, but some of them are time consuming especially when extracting a large number of samples. Here, we developed a rapid protocol for plasmid DNA extraction based on the alkaline lysis method of plasmid preparation (extraction at pH 8.0). Using this new method, a good plasmid preparation can be made in approximately one hour. The plasmids are suitable for any subsequent molecular applications in the laboratory. By applying the recommendations to avoid contaminations and to maximize the plasmid yield and quality during extraction, this protocol could be a valuable reference especially when analyzing a large number of samples.
Key words: Plasmid extraction, PCR, restriction enzymes, sequencing, contamination.
Simeon Oloni KOTCHONI, Eriola BETIKU and Olusola Olusoji SHONUKAN, Emma Wanjiru GACHOMO
Page: 1 - 10