DDRT PCR PDF

(a) Principle of the reactions underlying DDRT-PCR analysis. (N) Any of the four nucleotides G, A, T, or C. V can be any nucleotide besides T. (B) Any nucleotide. Recently, a new approach has successfully been developed: Differential-Display Reverse Transcription-PCR (DDRT-PCR). This technique has been proven to. Establishment of infection and disease implies modifications in the genetic programmes of the cell systems that are involved and the differential expression of.

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To investigate the upregulated or downregulated genes during the antagonistic process, both of the pathogenic and the antagonistic bacteria were grown either separately on two different nutrient agar plates or pce to each other on the same plate.

Differential display

Although chloramphenicol showed higher activity than our endophytic isolate; the antagonistic activity of this isolate is more preferred than this antibiotic. This claim is highly supported by the conditions which allow this enzyme not only to be produced but also to be over expressed.

Isolation, gene cloning and expression profile of a pathogen recognition protein: Acknowledgments We would like to thank King Khalid University, Faculty of Science, Biology Department for supporting this work with all the required chemicals and equipments.

Endophytes have been defined as pct bacteria or fungi which for all or part of their life cycle reside within the inner parts of plant tissues, but cause no symptoms of disease Vega et al. The sterilized cubic stem parts were then divided into two pieces using a sterilized razor blade. About ten bacterial colonies with the same colony shape and characteristics were obtained.

The up or down regulated genes were excised from the gel and submitted for purification using a gel purification kit Qiagene, Germany and the purified PCR products were submitted for sequencing Macrogen Inc. Conclusion The products of endophytic bacterial isolate B.

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Differential display – Wikipedia

The production of antibiotics and hydrolytic enzymes is a feature of many endophytic bacilli, including Bacillus cereus Pleban et al. Gene Ontology GO Terms. Either your web browser doesn’t support Javascript or it is currently turned off. However, we strongly propose that our isolate was able to produce serine protease for antimicrobial purpose.

Application of differential display RT-PCR revealed that the isolate was able to up-regulate a gene with serine protease like protein.

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The antagonistic effect was only observed with S. Khan Z, Doty SL. The antibacterial activity of the chosen bacterial isolate, named SA, was examined against S. After the incubation period, the S.

Both of sterile ddgt paper discs were loaded with the isolate SA and ten different antibiotic discs. Antagonistic effect of SA isolate against S. Multiple sequence alignment, molecular phylogeny and phylogenetic tree were performed using MEGA 5 software Fig. The protein ddrh well known as antimicrobial agent and was reported to be produced by plants, animals and insects.

The purified bacterium was checked for its ability to antagonize some human pathogenic bacteria including Klebsiella pneumoniaeEscherichia coli and S. A typical GC—MS chromatogram of the ethanol extracts inhibition zone slices.

In this study, Bacillus endophyticus strain SA isolated from the inner tissue of the stem of the cultivated plant Salvadora persica, Asir, Kingdom of Saudi Arabia produces an antagonistic factor. Phytochemical and pharmacological screening of Sterculiaceae species and isolation of antibacterial compounds.

To our knowledge, this is the first study that illustrates the bacterial production of serine protease for antagonistic function against another genus of bacteria.

Endophytic bacteria in Coffea arabica L.

Any external effect on the organism can affect its genetic behavior in a way of over or down expression of the genes according pfr the kind of the stimulant. In this study, Bacillus endophyticus strain SA isolated from the inner tissue of the stem of the cultivated plant Salvadora persicaAsir, Kingdom of Saudi Arabia produces an antagonistic factor.

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In addition, these microorganisms play an important role to protect their host plants from phytopathogenic fungi and bacteria Emmert and Handelsman The discovery of these affecting genes is related to the sequence of the primers used during the reverse transcription process. In-vivo expression technologies and differential display RT-PCR are providing new approaches to further examine a microbe’s response to experimental conditions which more closely resemble natural microbial associations and habitats.

The cultivated endophytic bacterial isolates were transferred to new nutrient agar NA plates for purification. This gene was up-regulated, excised from the gel and submitted for gene sequencing Fig. Identification of the producer strain was performed using the partial nucleotide sequence of 16S rRNA gene, which indicated that this strain is identical to B. Int J Pharm Pharm Sci. Molecular detection of the antagonistic factor using differential display RT-PCR technique To investigate the upregulated or downregulated genes during the antagonistic process, both of the pathogenic and the antagonistic bacteria were grown either separately on two different nutrient agar plates or close to each other on the same plate.

Serine protease has been produced by many organisms, including plants, animals and microbes Liu et al. To rate and determine the exact effect of the isolate SA against S. Impact of isolate SA activity in comparison with different antibiotics The antagonistic activity of the endophytic bacterial isolate was compared with ten different known bacterial antibiotics against S.