Phytases hydrolyse phytate (myo-inositol hexakisphosphate), an Anti-nutrition and the principal of phosphate stored in plant seeds to produce phosphate and lower phosphorylated myo-inositols.
The gene encoding holostable phytase from Nesterenkonia sp. strain F was replicated via PCR . The Escherichiacoli (DE3) and pET28b were used as the host and vector for cloning and expression of the phytase gene, respectively. Colony-PCR method and DNA sequencing is confirmed the cloning result.
Expression was induced by IPTG at a concentration of 0.5 mM. The molecular mass of the phytase was determined to be 48 KDa by SDS-PAGE.. the optimal PH and temperature of the enzyme were PH 6.5-7.5 and 20˚C , respectively. A higher enzyme activity was obtained when the gene expression was done in the presence of NaCl..
Due to the activity of the enzyme expressed in a wide range of temperatures, pH and salts, this enzyme will be widely used in a variety of industries in diverse conditions.

 Enzymes are biomolecules. They have the specific function on the particular substrate. Amylase constitute a class of industrial enzymes, representing approximately 30% of the world enzyme production. A gene encoding a halophilic α-amylase from Nestrenkonia sp.strain F was cloned. Nesterenkonia sp.strain F is a halophilic genus about the Micrococcacea family that contains high GC segments. The Escherichiacoli (DE3) and pET28b were used as the host and vector for cloning and expression of the α-amylase gene, respectively. Colony-PCR method and DNA sequencing is confirmed the cloning result.
The α-amylase gene was expressed in E.coli as a hexahistidine-tagged enzyme and purified. The molecular mass of the α-amylase was determined to be 52 KDa by SDS-PAGE. The optimal pH and temperature for enzyme activity were 7 and 45 ºC , respectively. It had a specific activity of 3.85 U/mg. The recombinant enzyme was active in a wide rang of salt concentration (0-4M) with it's maximum activity at 4M NaCl or 3.5M KCl. The enzyme was stimulated by Ca2+. It was strongly inhibited by Hg2+,but less affected by Cu2+, Fe2+, Zn2+ and Mg2+. These properties indicate wide potential applications of this recombinant enzyme in starch processing industries.
Also in the bioinformatics studies, the 3D structures of relate protein catalytic domain was investigated, by 3D structures modeling. 3D structure of α-amylase was composed of catalytic domain with (α/β) barrel fold.

Introduction: Scorpion venome is a mixture of active biological molecules with medical capability. Some of this peptide affecting on voltage gated sodium channels. This group usually has 60 to 70 amino acids, with 3 to 4 disulfide bonds. The ODNaTx8 gene coding a toxin that is a kind of beta-toxin sodium channel blocker that has similarity to a type of lipolysis activator factor and has been considered as a drug for lowering blood lipids, especially cholesterol. Material & Methods: The cDNA encoding Odontobuthus doriae sodium channel blocker toxins(ODNaTx8) was isolated from the library, replicated via PCR then these products and pET 28b extracted vector, double digested with restriction enzymes, then ligated and recombinant vector transferred to the E. coli BL21(DE3). Result: Observed the recombinant clones by transformation recombinant vectors to the E. coli BL21 and checked with clone-PCR and sequencing. For achieving recombinant protein, induced the recombinant clones with IPTG, then detected recombinant protein with SDS-PAGE technique. Conclutions: The importance of cloning the ODNaTx8 gene was to produce this toxin and purify it from a large variety of proteins. By purifying and identifying the product of the gene, it can be produced during the process of expression, producing an unlimited amount of its product, which is far more advantageous and less costly than taking a large number of animal samples for extracting their poisons and protein.

 Introduction: MMPs and TIMPs are involved in many cell signaling and Inflammatory processes and produced by many cell types, including astrocytes and microglial(glial) cells. Nonregulated MMP activity and an imbalance between metalloproteinases and their inhibitors might contribute to various disorders, including neurodegenerative diseases. Given their location and ubiquitous distribution, sialic acids (N-acetylneuraminic acid, NANA) can mediate or modulate a wide variety of physiological and pathological processes. So, evidence suggests that both MMPs and NANA play an important role in inflammatory neurodegenerative diseases. sialic acid involvement in the signaling pathways leading to MMP-9 and TIMP-1 gene expression in glial cells is unclear. Therefore, this study investigated the relationship between MMP-9, TIMP-1 and NANA and the effects of these ligand on the signaling processes of the neurodegeneration and inflammatory demyelination.
Materials and methods: Human Glial cell line was prepared from “Pasteur Institute of Iran” and cultured in DMEM, supplemented with 10% FBS. The IC50 value of NANA was obtained by MTT assay. Glial cell line was treated with NANA (300,500 and 1000 µg/ml) for 24h to investigate the effects of these ligand on the expression of MMP-9 and TIMP-1. Then total cellular RNA was isolated from approximately 10×106 Glial cells. 1000 ng of total RNA from each sample was used into cDNA. Real-time PCR determined the expression level of the MMP-9 and TIMP-1 transcripts and REST and SPSS software were used for analyze.
Results and conclusion: The results of MTT assay were analyzed by Excel software and IC50 was obtained equivalent to 1273.3 µM. Therefore, concentrations of sialic acid were selected for treatments that less than Ic50 (300, 500 and 1000 µM). By analyzing Real-time data, it was found that MMP-9 and TIMP-1 mRNA expression was up-regulated with treatment (300, 500 and 1000 µM). At concentrations of 300 and 500µM, there wasn’t significant difference between the up-regulated of two genes. It is concluded that along with the up-regulated of
MMP-9, TIMP-1 expression was also up-regulated. This event indicates cells attempt for maintaining MMP-9/TIMP-1 balance. MMPs expression is strictly controlled at different levels, to prevent their harmful effects and TIMPs are one of the MMPs controller. But it was found a significant difference between up-regulated of MMP-9 and up-regulated of TIMP-1, at 1000µM concentration of sialic acid. In this concentration, was observed imbalance at MMP-9/TIMP-1, similar to clinical reports of neurodegenerative diseases. These results indicate a possible involvement NANA on signaling pathway those genes expression.

Introduction: Multiple sclerosis (MS) is an inflammatory, demyelinating disease of the central nervous system(CNS). MS is determined by interaction between genes & environment. In the past decade, much attention has been given to vitamin D & its role in MS. Recent data implicate vitamin D in modulation of the risk as well as clinical course of disease. Since vitamin D acts through the vitamin D receptor (VDR) the aim of our study was to determine the association between the VDR rs731236 & rs7975232 polymorphisms & the MS risk in population of Khuzestan province.

Methods: This study comprised 150 MS cases , plus 150 healthy individuals as a control group. For all subjects, DNA testing for the VDR polymorphisms was done using PCR amplification and then RFLP and was confirmed by direct sequencing. Statistical analyses were performed using SPSS version 16.0.

Results: The rs731236 (A/G) polymorphism odds ratios for AA and GG genotypes were 1.2 (95% CI, 0.74 to 1.93; P=0.46) and 1.51 (95% CI, 0.66 to 3.46; P=0.33) compared with the AG genotype, respectively. The rs7975232 (C/A) polymorphism odds ratios for CA and AA genotypes were 8.65 (95% CI, 4.77 to 15.68; P=<0.001) and 1.38 (95% CI, 2.84 to 11.02; P=<0.001) compared with the CC genotype, respectively.

Conclusion: The rs7975232 polymorphism in VDR gene didn’t show association whit MS, but rs7975232 polymorphism showed good association whit this disease.
 

 phenytoin is one of the most important Chemical medicine used for the treatment of skin wounds. The aim of this study was to investigate the molecular function of phenytoin drug on gene VEGF and TGF-β expression. This study was accomplished on 30 male rats (15animals in each group) with approximately equal body mass. Skin wounds were made with an area of 2cm and a depth of 0.5 mm on the back of rats neck. Mice in each group divided to first week, second week and third week. Two groups were administered 2 times a day by topical phenytoin (1%) and Vaseline ( as control group(.Then gene expression was measured at days of 7، 14and 21. Results of the Real time PCR showed the increased expression of both VEGF and TGF-β in the first week and decreased expression in third week in the group receiving phenytoin compared with the control group. Expression changes were significant at the level of (p<0/001). Taking topical phenytoin (1%) increased the gene expression VEGF and TGF-β because these factors play a central role in wound healing، It can be stated that phenytoin done possibly its healing mechanism by stimulating the production of growth factors involved in wound healing.

 Enzymes are biomolecules. They have the specific function on the particular substrate. Enzymes
don’t have toxic effect on the environment. so they are very useful in the industrial processes.
Nesterenkonia sp.F is a halophilic genus about the Micrococcaceae family that contains high GC
segments.The RAST annotation pipeline software was used to annotate the DNA sequence.
Genome analyzing revealed presence of the, endo-1,4-beta xylanase gene produce
Endo-1,4-beta Xylanase Enzyme ( EC 3.2.1.8)as a hydrolyse enzyme which lyse endo 1,4-beta
glycosidic bond in the Xylan molecules of hemicellulose compounds. Here, a Xylanase gene
isolated from Nestrenkonia sp strain.F has been cloned in the pET26b expression vector. This
recombinant vector transformed in Escherichia.coli Dh5α and Esherichia.coli BL21(DE3). DNA
sequencing is confirmed the cloning results.
Also the computational calculations have revealed the possibilities of new endo-1,4-beta xylanase
functions. The functional properties of related proteins catalytic domain was investigated, by 3D
structures modeling. 3-D structure of endo- 1,4- beta xylanase was composed of catalytic domain
with (α/β) barrel fold. Results obtained docking studies have determined the binding sites for xylan
ligand and possible cofactors. The final information obviously indicated that despite differences in
amino acid sequence, the enzyme can recognizes xylan as a substrate.

 Introduction: Mitochondrial Trifunctional Protein(MTP) is a hetero-octamer compose of eight parts (subunits),4 α-subunits containing LCEH (long-chain 2,3-enoyl-CoA hydratase) and LCHAD (long-chain 3-hydroxyacyl CoA dehydrogenase) activity, and 4 b-subunits that encompass LCKT (long-chain 3-ketoacyl-CoA thiolase) activity which catalyzes three out of the β-oxidation spiral of long-chain fatty. This deficiency is an autosomal recessive disorder that causes a clinical spectrum of diseases. M-TFP deficiency are classified into 2 phenotypes: the more prevalent isolated LCHAD deficiency with defects of the α-subunits encoded by the HADHA (hydroxyacyl-CoA dehydrogenase α-subunit) gene and the less common pattern of complete M-TFP deficiency with defect both of HADHA and HADHB (hydroxyacyl-CoA dehydrogenase ß-subunit) genes.
Methods: A blood sample was collected from the patient and parent of the patient. isolation of DNA and Amplification of all exons of the HADHA and HADHB, directly sequence analyses of whole exons of both genes were performed.
Results and Conclusions: Results showed accumulation of long chain hydroxylated acylcarnitines. The molecular analysis of causative genes showed a novel homozygous missense mutation (p.Q385P) in exon 14 of HADHB gene. Since this mutation was not found in 50 normal control cases so It was concluded that mutation was a causative mutation. According to results this substitution may be alter the stability of enzyme structure because of altering total energy of structure. Bioinformatic analyzes confirmed these results.

Introduction: Infant respiratory distress syndrome is a acute pulmonary disease that the most common cause of respiratory distress in premature infants. lung is immature and non functional in this syndrome. Indeed, the surfactant deficiency causes the disease. Surfactant reduces the surface tension of the alveoli and thus prevents the collapse and atelectasis. ABCA3 lipid transporter protein plays an important role in the production of surfactant. The occurrence of mutations in this gene can affect the production of surfactant and the proper functioning of the lungs.
Materials & methods. In this study, the family that had a child with the disease was studied. The parents were clinically healthy. DNA was extracted from blood of all family members by Salting out method. To find the mutation (s), the results of the PCR of all patient's exons were analyzed by sequencing.
Results. According to the molecular genetic testing, a likely pathogenic (as softwares predicted) missense mutation G to A at codon 202 in exon 6 of the ABCA3 gene was found which alters the amino acid glycine to arginine. the patient was homozygous but the parents were heterozygous for this missense mutation at exon 6 . To confirm the pathogenic of the founded mutation, PCR and sequencing was performed for 50 normal controls.
Conclusion. According to the results and the absence of mutations in normal controls and predictions of various softwares, it can be concluded that the syndrome is probably caused by mutation in the Infant. Our findings show that the conversion of glycin to argentine mutation at codon 202, is new missense mutation in the population of the Southwest of Iran.
 

Introduction: Multiple sclerosis (MS) is an inflammatory, demyelinating disease of the central nervous system (CNS). Although the cause of MS is currently unknown, both genetic and environmental factors have been shown to contribute to the pathogenesis of MS. The human leukocyte antigen (HLA) class II alleles DRB1*1501,DRB5*0101, DQA1*0102, DQB1*0602 may have an important genetic effect. However, this is controversial in different population studies. Our aim was to investigate HLA DRB5*01 association with multiple sclerosis in some MS patients of Khuzestan population.
Methods: The present study focused on HLA DRB5*01 association in 202 MS patients and 187 healthy control from khuzestan. Female /male ratio was 4:1. HLA typing was performed by polymerase chain reaction (PCR) amplification with sequence-specific primers (PCR-SSP) method.

Results and Conclusions: 27.72% from patients and 21.39% from control group were positive with this type of HLA. This is the first study that investigate HLA DRB5*01frequency and association in multiple sclerosis patients in Khuzestan. We found that there is no association between HLA DRB5*01 with multiple sclerosis in Khuzestan province (P = 0.148)
 

Abstract:
Introduction: Multiple Sclerosis (MS) is a chronic, inflammatory demyelinating and neurodegenerative autoimmune disease of the central nervous system. In this disease, the immune system attacks myelin sheath around neurons. Despite extensive researches, the pathogenesis of this disease is still unknown, although there is evidence of complex interactions of both environmental and genetic factors. Recent evidence that CD8 T cells are implicated in MS suggests that HLA class I may also contribute. The negetive association and protective role of HLA-A*02 with multiple sclerosis has been confirmed. The aim of this study was to investigate the association of HLA–A*02 with multiple sclerosis in Khuzestan.
Materials and Methods: in this study 200 Khuzestan MS patients and 200 Khuzestan healthy controls were analysed. The male to female ratio was 1: 4. The majority (87.93%) of this patients presented with relapsing remitting MS (RRMS) and 12.07% of remaining were known as progressive relapsing (PR), primary progressive (PP) and secondary progressive (SP) MS. HLA typing was performed by polymerase chain reaction (PCR) amplification with sequence-specific primers (PCR-SSP) method.patient and healthy control that had HLA-A*02, showed two band on agarosis gel. The band size of HLA-A*02 was 249 bp and another band was 356bp and rlated to MOG gene (internal control).
Results: 29.5% of patients and 54% of healthy controls were positive with this HLA type.
Conclusion: According to this study HLA–A*02 genotype decreased the risk of MS in Khuzestan province.
 

Poly (ethylenimine) (PEI) is a cationic polymer commonly used in gene transfer/therapy protocols with high transfection efficiency both in vitro and in vivo. It is also cytotoxic, but the molecular basis of mechanism of its cytotoxicity is poorly understood. During this process PEI can interfere with apoptotic transcript in cell. Using transcriptom technique to identify the involved proteins is very helpful in this regard.
The aim of this research is the separation and recognition of the proapoptoses genes that connect to PEI and identification of the toxicity mechanism.
Method
Initially Human colon adenocarcinoma cells (HT29) were cultured in RPMI-1640 Media.Cells were homogenized by sonication and total RNA was isolated from control & transfected cell lines. RNA integrities were assessed by Nano-Drop(ND2000). cDNAwas synthesized using 2-step RT-PCR Kit.PCR arrey was performed using Real-Time PCR array Kit(cinnagen).Equal amounts of cDNAs were added to each PCR reaction.Transcrpted levels were normalized against transcript levels of endogenous control using ∆∆ct method.
Result
According the gene expression chartBclx,NfKB ,Bcl2 and Bclx genes expression significantly reduced in transfected HT29 cell lines while Bad,Bax, Casp3,9,7,8,cytoC ,Apaf-1 and TNFR1genes expression considerably increase in transfected HT29 cell lines compared to control cells.