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Aim and background: Cancer/testis (CT) antigens are a category of tumor antigens that are typically expressed only in the human germ line, and in several types of tumor. MAGEA4 is one member cancer testis antigen family that has relation with other tumors. Since the biological function of MAGEA4 is unclear, the aim of the present study was amplification and cloning of the gene in the expression vector to produce recombinant protein that expresses MAGEA4.
Material and method: Using PCR specific primers including restriction site, MAGEA4 was amplified. The purified PCR products were ligated between the BamH1 and Xho1 sites of pTZ57/R cloning vector and transformed into Escherichia coli Top10 strain and screened by IPTG and X-Gal. The correct orientation of MAGEA4 fragment was identified by restriction enzyme analysis and sequencing of constructed plasmid. Then sub cloning was carried out within pRUF expression vector with the same restriction site.
Result: The final confirmation was performed using colony PCR, double digesting and sequence analysis. Therefore the MAGEA4 gene was cloned into the restriction site of pRUF.
Conclusion: This study is an important step for producing recombinant proteins and is used to find the function of this gene for therapeutic targets.

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Aim and Background: Enterokinase is a gastrointestinal enzyme that acts as a protease with a specific sequence. Due to the ability of enterokinase to enzymatically digest the recombinant protein at its specific site (including the four amino acids Aspartic acid and one amino acid lysine), which is targeted at the protein, as a very useful tool for purification and separation Target protein from the non-target part is used in the pharmaceutical, food and industrial industries.
Materials and Methods: Independent variables used included OD (0.6, 1.2 and 1.8) at 600 nm, IPTG concentration (0.2, 0.5 and 0.8 mM) and The type of bacterial strain was (ShuffleT7, BL21, NICO21). The response surface methodology (RSM) in the form of a central composite scheme was used to predict independent variables on the production of enterokinase enzyme.
Results: OD equal to 1.8 at 600 nm wavelength, IPTG concentration of 0.71 mM and type of ShuffleT7 bacterial strain, LB culture medium, 2.5 mM lactose concentration and 25 ° C induction temperature, for production Recombinant enterokinase was optimized.
Discussion and Conclusion: Drug proteins play an important role in modern molecular medicine therapies. Enterokinase gene expression in the bacterial system facilitates the purification of low concentrations, so large-scale, highly purified recombinant proteins can be produced and optimized in the bacterial system.

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Aim and Background: One of the most malignant and common brain tumors is glioblastoma multiforme (GBM). Patient survival is relatively poor, and due to the difficulty of early diagnosis, most patients die within one year of diagnosis. The aim of this study was to investigate the expression of YKL39 as a possible biomarker in glioblastoma multiforme and to evaluate the miR of this gene.

Materials and Methods: In this study, 25 tissue samples with glioblastoma multiforme before treatment and 25 samples of healthy tissue with glioblastoma multiforme tumor were collected as a control sample. RNA extraction and cDNA synthesis were performed. YKL39 gene expression was assessed using real-time PCR and ACTB gene was used as internal control. Statistical analysis of data was performed using GraphPad Prism software version 8. The Roc curve was used to evaluate the biomarker value of YKL39 gene.

Results: High expression of YKL39 (P **** <0.0001) was observed in glioblastoma tissue samples. Expression showed that YKL39 gene was not statistically significant with the age of patients with P value = 0.2971 and gender with P value = 0.888.

Conclusion: The increase in YKL39 gene expression in glioblastoma tissue samples is significant compared to healthy tumor margin tissue. The Roc curve showed that the YKL39 gene could not have biomarker value.
 

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Aim and Background: glioblastoma Multiforme (GBM), astrocytoma grade IV which makes up 53% of all gliomas. It is the most common and deadly primary malignant brain tumor in adults. The mortality rate in glioblastoma multiforme is very high. The aim of this study was to determine the expression of SLC1A2 gene in tumor tissues and margins Healthy subjects with glioblastoma multiform were designed and performed.
Materials and Methods: In this study, 50 tissue samples including 25 tissue samples with glioblastoma multiform disease and 25 healthy tissue samples around the GBM tumor before treatment were collected as control samples. RNA extraction and cDNA synthesis were performed in all samples. SLC1A2 gene expression was investigated using Real-time PCR method. ACTB gene was used as internal control. Statistical analysis of data was done using GraphPad Prism version 8 software. Roc curve was used to check the biomarker value of SLC1A2 gene.
Results: Decreased SLC1A2 gene expression was observed in glioblastoma multiforme samples and its expression was compared with the age, gender and tumor size of the patients. The expression of SLC1A2 gene with the age of the patients was statistically investigated and a significant relationship was observed among people over 50 years old and under 50 years old (P-value˂ 0.05), while this significant relationship was not observed in gender and tumor size (P-value>0.05).
Conclusion: The SLC1A2 gene was less expressed in the tumor tissue of patients with glioblastoma multiforme than in the healthy tissue around the tumor. By examining the ROC curve, it is possible that the SLC1A2 gene can be proposed as a biomarker.