فصلنامه تازه های بیوتکنولوژی سلولی مولکولی
پیاپی 36 (پاییز 1398)

Escherichia coli is the first bacteria on which cloning and protein expression are carried out. The purpose of this study was to summarize the numerous articles published in the field of protein expression in Escherichia coli. In studies, researchers in a new project require the production of pure protein, which at the theory level is easy to obtain a recombinant protein, but   there are virtually many problems, including low growth of host, formation of inclusion, protein not being active, and there is even no protein production in the exprimental scale. To enhance the expression of the protein, factors such as suitable host, vector design, transcriptional settings, selective markers, and reluctant labels are effective. There are a range of fusion proteins that can effect on accurate folding, solubility, and proteolytic resistance. Also the ability to target produced proteins to cytoplasm, periplasm, membranes and the culture medium, and some techniques grant the ability of post translational modifications in prokaryotic cells. Therefore, the key to the successful expression of recombinant proteins in E. coli is a combination of expert manipulation of the components of a broad genetic apparatus.

CD133 is a membrane glycoprotein that contains 865 amino acids and is normally expressed in hematopoietic stem cells, endothelial precursor cells, neuronal and glial stem cells. The CD133 marker has been used to identify cancer cells and many malignancies, including the brain, colon and lung. The aim of this study was to evaluate the expression level of CD133 as a marker for the detection of leukemia.

Blood samples were collected from 30 patients with leukemia and 30 healthy controls. After RNA extraction, cDNA was synthesized and CD133 gene expression was evaluated using Real-Time PCR technique. Data were analyzed using SPSS 16.0 software.

According to the obtained data and statistical analysis (p <0.05) using SPSS software, the expression level of CD133 gene in cancerous samples was significantly increased compared to healthy ones. That (p CD133 = 0.0065) was obtained.
Discussion and

Given the role of CD133 as a marker for the identification of many cancers, it can be concluded that CD133 can be a good marker for cancer detection and prevention. Be leukemia

Nanoparticles are widely used in medical, pharmaceutical and health sciences. The aim of this project was to evaluate the mesophilic bacteria isolated from the Gulf coast in the production of silver nanoparticles and investigate the antimicrobial effect of these nanoparticles on some pathogenic bacteria.

This descriptive and cross-sectional study was performed over a period of 8-month from April to November 2017. The isolates were purified from water and sediments samples of the coasts of Hormozgan Province - Iran, after that the purified isolates were cultivated in Zobell Marine Broth medium. The obtained supernatant of the medium was added to the silver nitrate (AgNO3) solution (0/001 M) with (1:5) ratio at the light condition in order to the reduction of AgNO3 to metallic silver. The synthesis of silver nanoparticles (AgNPs) was investigated by UV-Vis spectrophotometer, Transmission electron microscope(TEM), and Fourier transform infrared (FTIR) spectroscopy analysis. The antimicrobial activity of AgNPs was evaluated against 6 microbes.

The results indicated that supernatants of all isolates are capable of producing silver nanoparticles. The surface plasmon resonance of AgNPs showed a maximum peak near 420 nm, which UV–vis spectra correspond to the absorbance of AgNPs. The results of TEM micrographs image of silver nanoparticles produced by dominant strain showed spherical shapes and size of 2/88 to 19 nm. The synthesized AgNPs showed the antimicrobial activity and inhibitory effects on the growth of tested microbes. 

The desired isolates have a potential ability to producing AgNPs, and to summarize, this is a low cost, ecofriendly, and quick method for the synthesis of AgNPs.

Bacterial infections of the genital system are common causes of infertility. One of the most important causes of male infertility is seminal and genital tract infections. In the meantime, a wide range of bacteria, in varying degrees, contribute to infertility in men. Helicobacter pylori may be involve in infertility. Because antibodies against this bacteria have been significantly detected in the genital fluids of infertile patients. The aim of this study was to determine the presence of Helicobacter pylori in the semen of the infertile men with rapid molecular PCR method and determine its percentage.

100 samples of semen collected from Saram hospital and DNA Extracted from these specimens using Boiling/DNG_PLUS method.  Optimized PCR test was performed on samples. PCR test evaluated from the respect sensitivity and specificity.

In this study, amplicon with the size of 294 bp, observed with agarose electrophoresis.  In the specificity test, primers created only a band for H.pylori DNA. Of the 100 samples tested, only 10 samples (10%) were positive for DNA of H.pylori.

According to the results of this study, H.pylori may be one of the bacterial agents that can be considered for male infertility, of course requires further studies. While PCR is suitable, quick and sensitive molecular technique for detection of H.pylori in infertile men.

From the perspective of biology  The genus Leishmania belongs to Tripanozomatite tribe flagellate Parasits. Leishmania is in the form of promastigotes and amastigotes. In culture, In the form of promastigotes and intra-macrophage is in the form of amastigotes. The aim of this study was to investigate the survival of L.tropica parasites in culture .

In this study, the Possibility of growth in NNN and RPMI1640  culture  Reviewed . In the laboratory, Conditions were provided that  L.tropica  protighotites  transformed into amastigotes that live in the host cell. Growth of L.tropica  was investigated in comparison with other types of leishmaniasis, such as L. major. L.tropica was kept at 25-26 ° C in a conventional incubator. L.tropica culture culture was performed in RPMI1640 culture. Contamination of L.tropica was done on J774A-1 cells. The J774A-1 cell was maintained in RPMI1640 medium. Cells were stored at 37 ° C in a CO2-encoded incubator.

L.tropica  After infecting  was observed on the J774A-1 macrophage cell line. compared to L.major, Which takes more time to show its performance, L.tropica  is the power of survival for a long time Inside it . It was also observed that L.tropica was only stored and not proliferated in NNN medium.

 The results indicate that L.tropica  protighotites, in laboratory environment Became amastigotes Which host cells have the power of survival for a long time.

 In recent years, science and industry have focused on preparing nanoparticles based on the principles of green chemistry. For this purpose, various types of biological structures such as bacteria, yeasts, and string molds are used. Due to the resistance of bacteria to antibiotics, the need to replace effective antimicrobials, with fewer side effects is less. The aim of this research was to synthesis the iron oxide nanoparticles by lactobacillus fermentum cytoplasmic extract and investigate its antimicrobial effects against Staphylococcus aureus and Pseudomonas aeruginosa.

In this study, after preparing cytoplasmic extract of Lactobacillus fermentum from frees-thow method, iron sulfate solution 10-3 M were added and incubated for 3 weeks in the presence of 5 % carbon dioxide. Production of nanoparticles was investigated by X-ray Diffraction (XRD) and Transmission Electron Microscopy (TEM) and finally, the antimicrobial effects of standard strains of two bacteria (Staphylococcus aurous and Pseudomonas aeruginosa) were determined using agar diffusion method.

Changing the color of solution to black is an indication of iron oxide nanoparticles production. The formation of iron oxide nano-crystals by Lactobacillus fermentum cytoplasmic extract was shown by XRD analysis and the average nanoparticles sizes that determined by transmission electron microscopy TEM were found to be about 10-15 nm with a spherical shape. The mean diameter of the no-growth field indicates that the synthesized nanoparticles inhibit Staphylococcus aureus in concentrations of 100 and 1000 mg/ ml, while are ineffective against  Psedumonas aeruginosa.

The use of cytoplasmic extract of Lactobacillus fermentum can be introduced as an effective biological method for the production of iron oxide nanoparticles and with more studies in this area, green nanoparticles may be used as suitable candidates for the treatment of microbial infections.

The purpose of this study was to investigate the association between different polymorphisms of the vitamin D receptor coding gene (Taq1, Bsm1, Apa1, Fok1) with renal disease in patients with type 2 diabetes.

In this case-control study, blood samples were taken from 50 healthy controls and 50 type 2 diabetic patients as control and 50 type 2 diabetic patients with nephropathy. Then, SSP-PCR technique was used to determine the different genotypes of vitamin D receptor polymorphisms and the results were analyzed by SPSS 22 software.

In type 2 diabetic patients with Bb (64%) genotypes were the most frequent genotypes for Bsm1 polymorphism, FF (54%) for Fok1, Aa (84%) for Apa1 and TT (36%) for Taq1. In addition, the probability of Nephropathy with bb genotype (P-Value = 0.007, OR = 6.7730) for Bsm1 polymorphism, ff genotype (OR = infinity and P-Value = 0.01) for Fok1 polymorphism, Aa genotype (OR) = 5.25 and P-Value <0.001) for Apa1 polymorphism and tt genotype (OR = 5.41, P-Value = 0.003) for Taq1 polymorphism increased.

Genotypic frequency of Bsm1, Apa1, Fok1 and Taq1 polymorphisms is related to the incidence of renal disease (Nephropathy) in type 2 diabetic patients. However, the correlation between different polymorphisms and the incidence of kidney disease shows that Apa1 polymorphism has the greatest effect on renal complications in patients with type 2 diabetes. In addition, Bsm1 polymorphism has the highest effect on the incidence of type 2 diabetes.

Bacterial infections of the genital system are common causes of infertility. One of the most important causes of male infertility is seminal and genital tract infections. In the meantime, a wide range of bacteria, in varying degrees, contribute to infertility in men. Helicobacter pylori may be involve in infertility. Because antibodies against this bacteria have been significantly detected in the genital fluids of infertile patients. The aim of this study was to determine the presence of Helicobacter pylori in the semen of the infertile men with rapid molecular PCR method and determine its percentage.

100 samples of semen collected from Saram hospital and DNA Extracted from these specimens using Boiling/DNG_PLUS method.  Optimized PCR test was performed on samples. PCR test evaluated from the respect sensitivity and specificity.

In this study, amplicon with the size of 294 bp, observed with agarose electrophoresis.  In the specificity test, primers created only a band for H.pylori DNA. Of the 100 samples tested, only 10 samples (10%) were positive for DNA of H.pylori.

According to the results of this study, H.pylori may be one of the bacterial agents that can be considered for male infertility, of course requires further studies. While PCR is suitable, quick and sensitive molecular technique for detection of H.pylori in infertile men.