فهرست مطالب غزل قجری

Compared to other primary photosynthetic products (such as sucrose and starch), little is known about sugar metabolism and how it is integrated with others. Mannose-6-phosphate reductase (M6PR) is a key enzyme involved in mannitol biosynthesis in celery. This study aimed to clone the gene, express and purify the M6PR enzyme and investigate its function on mutated genes in a laboratory environment.
 
First, the mRNA was extracted from the celery plant, then the cDNA was synthesized, and the product was used as a template to amplify the M6PR gene. The PCR product was purified in a DNA gel extraction kit. The purified PCR product was cloned into the pTZ57R vector according to the T/A Cloning recipe (Fermentase Company). Susceptible cells of E.coli strain Top10 were prepared using the biochemical method of calcium chloride, and the recombinant vector inside it was transformed and cultured on a plate containing ampicillin. The cloning accuracy was done using M6PR gene PCR and enzymatic digestion of the recombinant plasmid by BamHI and SacI enzymes (Fermentase Company). The M6PR gene was homogenized in the expression plasmid pET32a and transferred into E.coli strain BL2I. The promoter was induced with IPTG and analyzed by western blotting. The protein was purified by affinity chromatography column (His. Tag/S.Tag).
The results showed that the enzyme could identify the heteroduplex regions of the gene. The recombinant M6PR purified from Escherichia coli had specific molecular activity. The results of double digestion of the plasmid with SacI and BamHI enzymes were 2870 bp and 186 bp fragments. According to the blast test result, the current fragment had 100% similarity with the M6PR gene of the celery plant. M6PR recombinant gene transcription results showed that the M6PR recombinant gene transcription rate was 2.3 in the transgenic strains and 0.32 in control, which showed a statistically significant difference at the P<0.01 level. After induction of the promoter and sampling at different times, the samples on the SDS-PAGE gel showed a protein band in the region of 42 kDa, indicating the protein's successful expression.
 
Homology of M6PR enzyme gene obtained from celery plant and then recombinant production of this enzyme in the laboratory can lead to its high expression in the prokaryotic system so that the enzyme has activity. Also, the present study showed that plant enzymes are active when expressed in bacteria and can be used as a suitable source to accelerate catabolic activity.

Acinetobacter baumannii (A. baumannii) is a non-fermentative gram-negative coccobacillus that shows high resistance to antimicrobial compounds. Biofilm formation is one of the essential features of many Acinetobacter species that leads to high antibiotic resistance. This study aimed to evaluate the ability of biofilm formation and determine the antibiofilm activity of chitosan nanoparticles in clinical isolates of A. baumannii.

This descriptive cross-sectional study was conducted in 2021 and investigated 100 isolates collected from different hospitals. Microscopic, biochemical, and molecular tests were performed to identify the bacteria. The antibiotic resistance pattern of the isolates was evaluated by the disk diffusion method against 10 antibiotics, and the ability to produce biofilm was evaluated by microtiter plate method. Subsequently, 16SrRNA and CsuA genes were identified by multiplex-PCR molecular methods. After the preparation of chitosan nanoparticles and determination of MIC concentration, antibiofilm activity was measured by plate microtiter, and Real-Time PCR was used to examine the expression of the CsuA gene involved in biofilm.

In this study, out of 100 isolates examined, 29 isolates were confirmed as A. baumannii. Among 29 isolates, ceftazidime had the highest drug resistance (75.86%). The CsuA gene was detected in 51.72% of the isolates. Moreover, using a microtiter plate and Real-Time PCR, the level of antibiotic activity of chitosan nanoparticles was determined at a significant level of P<0.01.

Considering the anti-biofilm effects found in the present study, it seems that chitosan nanoparticles can be used as a pharmaceutical candidate in the pharmaceutical industry.

Oilseeds are the second-largest source of calories for human societies after cereals. Sesamum indicum has been considered recently due to its oil quality and content of 43-46% unsaturated fatty acids. To investigate the possibility of transferring the recombinant aroA-CYP81Q1 gene through Agrobacterium tumefaciens to the Sesamum indicum plant (reported for the first time in the country), after optimizing tissue culture and selecting the best hormonal combination, a factorial experiment was performed. 

In this experiment, the recombinant aroA-CYP81Q1 gene was synthesized and transformed into an Agrobacterium strain (LB4404). Gene cloning was confirmed by PCR and then confirmed by sequencing. The efficacy and frequency of transgenic Sesamum indicum were evaluated in the selected medium containing 50 mg/l kanamycin. Finally, to confirm the transgenicity of the regenerated plants, PCR analyzes were performed with specific primers on selected plants. Also, the expression of the sesamin-producing gene (CYP81Q1) in control groups and transgenic groups was measured at a significant level of P <0.01.   

Statistical analysis showed that the percentage of regeneration of transgenic seedlings using Agrobacterium (LB4404) in the selected medium containing kanamycin was 33%. Also, PCR analysis of transgenic plants showed a prevalence of transgenics of 33%. In addition, amplification of the bp fragment indicated the accuracy of cloning in transgenic plants. The results indicate the successful transfer of this gene to the sesame plant to increase its industrial properties. The results of the analysis of variance showed that there was a significant difference in the expression of sesamin-producing genes between transgenic cultivar and control groups (P <0.01). 

Since pBI121-based expression vectors are more efficient than other expression vectors and are widely used in the transfer of recombinant genes to plants, the recombinant vector constructed in this study indicates successful gene transfer. Sesame to sesame plant to increase its industrial properties.