(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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Bacillus subtilis BS as an antagonist of potato blackleg and soft rot bacteria. Antibacterial activity of volatile component and various extracts of Spirulina platensis. However, only one of the sequenced genes was matched with genes deposited in GenBank.
Differential display – Wikipedia
Differential display using Ea1 and Ea2 primers combined with OP1 reverse transcription primer. These results confirm the common concept regarding fdrt ability of microorganism to develop new strategies to resist more and more antibiotics, which invite the researchers to find new and effective antimicrobial agents. On the other hand, only chloramphenicol recorded 2. Both of sterile filter paper discs were loaded with the isolate SA and ten different antibiotic discs.
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. The antagonistic factor was isolated from the bacterial culture medium and purified by thin layer chromatography technique, then analyzed by GC-MS analysis. Chitinolytic activity of an endophytic strain of Bacillus cereus. In addition, these microorganisms play an important role to protect their host plants from phytopathogenic fungi and bacteria Emmert and Handelsman Phytochemical pxr pharmacological screening of Sterculiaceae species and isolation of antibacterial compounds.
Differential display can offer some favors regarding this point, especially in the identification of some up-regulated or down-regulated genes under specific conditions. Antagonistic bioactivity of an endophytic bacterium isolated from Epimedium brevicornu Maxim. A review on production of serine alkaline protease by Bacillus spp. Comparing the antagonistic effect of the isolated bacteria SA with different antibiotics To rate and determine the exact effect of the isolate SA against S.
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The discovery of these affecting genes is related to the sequence of the primers used during the reverse transcription process. It is widely produced by Bacillus species and found tremendous applications in pharmaceuticals Bhunia et al.
The reasons for this hypothesis came from that there are many recorded disadvantages of chloramphenicol including the risk of its side effects, rising of resistance rates and uncomfortable posology Diniz-Santos et al. The extraction of the active metabolites produced by the endophytic isolate SA was done as follows: Unveiling of up-regulated or down-regulated genes Any external effect on the organism can affect its genetic behavior in a way of over or down expression of the genes according to the kind of the stimulant.
The protein is well known as antimicrobial agent and was reported to be produced by plants, animals and insects. 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.
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Serine protease is also known to be produced by bacteria for purposes oth er than bacterial—bacterial antagonistic effect, which has been confirmed by this study.
Conclusion The products of endophytic bacterial isolate B.
However, we strongly propose that our isolate was able to produce serine protease for antimicrobial purpose. Biocontrol of plant disease: The mixture was filtered using filter papers to remove the agar medium pieces and the ethanolic dcrt was undergoing to GC—MS analysis as described previously Ezhilan and Neelamegam ; Moustafa et al.
Detected compounds and its derivatives showed various biological activities, for example, Ganendren et al. Identification of the producer strain was ddet using the partial nucleotide sequence of 16S rRNA gene, which indicated that this strain is identical to B.
We would like to thank King Khalid University, Faculty of Science, Biology Department for supporting this work with all the required chemicals and equipments. The number and extent of the genes affected is not totally known to others. Serine protease is also known to be produced by bacteria for purposes oth er than bacterial-bacterial antagonistic effect, which has been confirmed by this study. 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.
About ten bacterial colonies with the same colony shape and characteristics were obtained. Serine protease is an enzyme that produced by many organisms including bacteria. The antagonistic factor was isolated from the bacterial culture medium and purified by thin layer chromatography technique, then analyzed by GC—MS analysis. Braz J Infect Dis. The Bacillus subtilis genome sequence: Application of differential display RT-PCR revealed that the isolate was able to up-regulate a gene with serine protease like protein.
The bacterial production and secretion of serine protease for antimicrobial function is not yet documented. Serine proteases from nematode and protozoan parasites: