Targeted Deletion of Los1 Homologue Affects the Production of a Recombinant Model Protein in Pichia pastoris     

Najmeh Zarei  Hosnieh Ghasemi  Mahsa Nayebhashemi  Mozhgan Zahmatkesh  Monire Jamalkhah  Nafiseh Moeinian  Zahra Mohammadi  Somayeh Enayati  Vahid Khalaj 《Iranian Biomedical Journal》2021,25(4):255

Background:The methylotrophic yeast Pichia pastoris is an appealing production host for a variety of recombinant proteins, including biologics. In this sense, various genetic- and non-genetic-based techniques have been implemented to improve the production efficiency of this expression platform. Los1 (loss of supression) encodes a non-essential nuclear tRNA exporter in Saccharomyces cerevisiae, which its deletion extends RLS. Herein, a los1-deficient strain of P. pastoris was generated and characterized. Methods:A gene disruption cassette was prepared and transformed into an anti-CD22-expressing strain of P. pastoris. A δ los1 mutant was isolated and confirmed. The drug sensitivity of the mutant was also assessed. The growth pattern and the level of anti-CD22 ScFv expression were compared between the parent and mutant strains.Results:The los1 homologue was found to be a non-essential gene in P. pastoris. Furthermore, the susceptibility of los1 deletion strain to protein synthesis inhibitors was altered. This strain showed an approximately 1.85-fold increase in the extracellular level of anti-CD22 scFv (p < 0.05). The maximum concentrations of total proteins secreted by δ los1 and parent strains were 125 mg/L and 68 mg/L, respectively.Conclusion:The presented data suggest that the targeted disruption of los1 homologue in P. pastoris can result in a higher expression level of our target protein. Findings of this study may improve the current strategies used in optimizing the productivity of recombinant P. pastoris strains. Key Words: Aging, Longevity, Pichia pastoris, Recombinant proteins  相似文献   

Transplanting P75-Suppressed Bone Marrow Stromal Cells Promotes Functional Behavior in a Rat Model of Spinal Cord Injury     

Houri Edalat  Zahra Hajebrahimi  Vahid Pirhajati  Mansoureh Movahedin  Mahmoud Tavallaei  Mohammad-Reza Soroush  Seyed Javad Mowla 《Iranian Biomedical Journal》2013,17(3):140-145

Background: Bone marrow stromal cells (BMSC) have been successfully employed for movement deficit recovery in spinal cord injury (SCI) rat models. One of the unsettled problems in cell transplantation is the relative high proportion of cell death, specifically after neural differentiation. According to our previous studies, p75 receptor, known as the death receptor, is only expressed in BMSC in a time window of 6-12 hours following neural induction. Moreover, we have recently reported a decreased level of apoptosis in p75-suppressed BMSC in vitro. Therefore, our objective in this research was to explore the functional effects of transplanting p75:siRNA expressing BMSC in SCI rats. Methods: Laminectomy was performed at L1 vertebra level to expose spinal cord for contusion using weight-drop method. PBS-treated SCI rats (group one) were used as negative controls, in which cavitations were observed 10 weeks after SCI. pRNA-U6.1/Hygro- (group two, as a mock) and pRNA-U6.1/Hygro-p75 shRNA- (group three) transfected BMSC were labeled with a fluorescent dye, CM-DiI, and grafted into the lesion site 7 days after surgery. The Basso-Beattie-Bresnehan locomotor rating scale was performed weekly for 10 weeks. Results: There was a significant difference (P≤0.05) between all groups of treated rats regarding functional recovery. Specifically, the discrepancy among p75 siRNA and mock-transfected BMSC was statistically significant. P75 siRNA BMSC also revealed a higher level of in vivo survival compared to the mock BMSC. Conclusion: Our data suggest that genetically modified BMSC that express p75:siRNA could be a more suitable source of cells for treatment of SCI. Key Words: Spinal cord injury (SCI), Apoptosis, Bone marrow stromal cells (BMSC)  相似文献   

Changes of vitamins A, E, and C and lipid peroxidation status of breeding and pregnant sheep during dry seasons on medium-to-low quality forages     

Mohebbi-Fani M  Mirzaei A  Nazifi S  Shabbooie Z 《Tropical animal health and production》2012,44(2):259-265

This study assessed the changes of plasma vitamin A, E, and C and the lipid peroxidation status of sheep during breeding and pregnancy under drought conditions. The study was conducted on 105 cross-bred fat tailed ewes, 3–5 years old with body condition scores (BCS) of 2.5 to 3.5. The ewes were grazing on medium-to-low quality forages during summer and low quality forages within the succeeding months and had ad libitum access to a mixture of alfalfa hay (40%) and wheat straw (60%) in the afternoons. From 3 weeks before breeding till 1 month after the introduction of rams, 300 g of barley grain/head/day was offered to the ewes and then the supplemental grain was reduced to 100 g/head/day. For better synchronization of estrus cycles in ewes, they were isolated from the rams for at least 2 months and then kept in close proximity of the rams for 1 week before the introduction of the rams to the ewe flock. Then, whole blood samples were collected on days 1, 7, 21, and 120 after ram introduction. Vitamins A, E, and C were measured in plasma. Malondialdehyde (MDA) was measured in the hemolysate as a biomarker of lipid peroxidation. Plasma progesterone (P4) was measured in the samples of day 120 for assessing pregnancy status of the ewes. Vitamins A and C showed continuous and significant declines (P < 0.05) through days 1 to 120. Vitamin E declined only during the first 21 days of the study and remained almost constant till day 120. MDA concentration increased significantly at day 21. An elevated concentration of MDA was also detected at day 120. The difference between days 21 and 120 was not significant (P > 0.05). A positive correlation between vitamins E and C was detected at day 120 (r = 0.349, P < 0.01). Age and BCS did not affect the patterns of changes. Assuming that the ewes with P4 concentrations ≥2.5 ng/ml were pregnant, 95 out of 105 ewes (90.5%) were pregnant at day 120 of the study. Under the conditions of the present study with medium-to-low quality pastures as the main sources of feed, ewes of various ages and body conditions may suffer from oxidative stress during breeding and pregnancy.  相似文献   

Fumigant toxicity of Carum copticum and Vitex pseudo-negundo essential oils against eggs, larvae and adults of Callosobruchus maculatus     

Bibi Zahra Sahaf  Saeid Moharramipour 《Journal of pest science》2008,81(4):213-220

Fumigant activity of essential oil vapors distilled from Carum copticum C. B. Clarke and Vitex pseudo-negundo Hand I. MZT. was tested against eggs, larvae and adults of Callosobruchus maculatus (F.). Fumigant toxicity was assessed at 27 ± 1°C and 60 ± 5% RH, in dark condition. The influence of different concentrations of the essential oil vapors on egg hatchability, larval and adult mortality was significant. Data probit analysis showed that lethal concentration of the essential oil to kill 50% of the population (LC₅₀) for egg, larvae and adult were found to be 1.01, 2.50 and 0.90 μl/l air of C. copticum oil, followed by 2.20, 8.42 and 9.39 μl/l air essential oil of V. pseudo-negundo, respectively. Between these essential oils, C. copticum was almost more toxic than V. pseudo-negundo on all growth stages of C. maculatus. The present study suggests that essential oils from these medicinal plants may be potential grain protectants as botanical alternative fumigants and could be used in the management of various life stages of C. maculatus.  相似文献   

The selection of superior walnut (<Emphasis Type="Italic">Juglans regia</Emphasis> L.) genotypes as revealed by morphological characterization     

Zahra Rezaei  Ali Khadivi  Babak ValizadehKaji  Ahmadreza Abbasifar 《Euphytica》2018,214(4):69

Walnut (Juglans regia L.), is one of the main economic fruit species in the world and is rich in nutritious substances. In order to obtain superior walnut genotypes, a set of seedling walnut genotypes were evaluated and their results were recorded. The range of nut length was between 25.57 and 47.77 mm. Also, the ranges of nut weight, kernel weight and kernel percentage varied from 5.35 to 21.31 g, 2.49 to 11.15 g and 37.27 to 66.29%, respectively. Simple correlation analysis showed significant positive and negative correlations between some of the measured traits. Principal component analysis reduced the evaluated traits into the seven main components, which explained 71.71% of the total variance. Bi-plot analysis performed using the first two components, distributed the genotypes into four sides of the plot. Based on the present results, a high variation was observed among the trees, indicating a high potential of the studied germplasm to select the superior genotypes. According to the present research, 57 genotypes with having the highest nut weight and kernel weight, as well as bright color of the kernel, were recognized as superior genotypes and can be used to establish the new orchards or improve cultivars.  相似文献   

Village Common Forest Management in Komolchori,Chittagong Hill Tracts,Bangladesh: An Example of Community Based Natural Resources Management     

Md. Arif Chowdhury  Fatima-Tuz- Zahra  Md. Farhadur Rahman  Kamrul Islam 《Small-Scale Forestry》2018,17(4):535-553

Chittagong Hill Tracts (CHTs) in Bangladesh covers about 50% of the country’s natural forest resources, providing sources of food, fuel-wood, medicinal plants, timber and water supply to the ethnic communities living in the area. The ethnic communities in CHTs have been managing a small patch of forest known as a village common forest (VCF) adjacent to their village following a community based forest management approach. Taking a case study of Komolchori VCF, this study examined dependency of community people on VCF resources, their conservation-related attitudes and awareness, indigenous management, and forest phyto-sociological structure. A survey and focus group discussions were conducted among the settlers and non-settlers in Thana Chandra para and Jaduram para villages of the Komolchori VCF. Key informant interviews were conducted with two non-government organizations (NGOs) and the headman in the study area. To assess phyto-sociological structure of vegetation, 25 quadrats (10 m?×?10 m) were laid out randomly. Results revealed that non-settlers were more inclined towards VCF conservation and women played a crucial role as they were directly involved in forest products and water collection. Soil erosion caused by shifting cultivation has made community people more aware of forest degradation. Training program provided by NGOs on different horticultural techniques and a “Saving-Credit program” improved livelihood conditions and reduced pressures on VCF resources. Locally devised traditional rules guide the community people to manage the VCF sustainably. The vegetation survey showed that the density of Komolchori VCF was 1420 trees/ha with a basal area of 27.98 m²/ha. The Shannon–Weiner index, species diversity index and index of dominance in the study area were 2.91, 0.62, 0.09, respectively.  相似文献   

Modelling the impact of climate change on rangeland forage production using a generalized regression neural network: a case study in Isfahan Province,Central Iran     

JABERALANSAR Zahra 《干旱区科学》2017,9(4):489-503

Monitoring of rangeland forage production at specified spatial and temporal scales is necessary for grazing management and also for implementation of rehabilitation projects in rangelands. This study focused on the capability of a generalized regression neural network(GRNN) model combined with GIS techniques to explore the impact of climate change on rangeland forage production. Specifically, a dataset of 115 monitored records of forage production were collected from 16 rangeland sites during the period 1998–2007 in Isfahan Province, Central Iran. Neural network models were designed using the monitored forage production values and available environmental data(including climate and topography data), and the performance of each network model was assessed using the mean estimation error(MEE), model efficiency factor(MEF), and correlation coefficient(r). The best neural network model was then selected and further applied to predict the forage production of rangelands in the future(in 2030 and 2080) under A1 B climate change scenario using Hadley Centre coupled model. The present and future forage production maps were also produced. Rangeland forage production exhibited strong correlations with environmental factors, such as slope, elevation, aspect and annual temperature. The present forage production in the study area varied from 25.6 to 574.1 kg/hm~2. Under climate change scenario, the annual temperature was predicted to increase and the annual precipitation was predicted to decrease. The prediction maps of forage production in the future indicated that the area with low level of forage production(0–100 kg/hm~2) will increase while the areas with moderate, moderately high and high levels of forage production(≥100 kg/hm~2) will decrease both in 2030 and in 2080, which may be attributable to the increasing annual temperature and decreasing annual precipitation. It was predicted that forage production of rangelands will decrease in the next couple of decades, especially in the western and southern parts of Isfahan Province. These changes are more pronounced in elevations between 2200 and 2900 m. Therefore, rangeland managers have to cope with these changes by holistic management approaches through mitigation and human adaptations.  相似文献   

Assessment of the Allelopathic Potential of Cumin Accessions in Different Soil Water Potential     

Zahra Ghafari  Hassan Karimmojeni  Mohammad M. Majidi  Bahare Naderi 《Journal of Crop Science and Biotechnology》2018,21(3):249-260

Allelopathy is one effective solution to reach a healthy life without any pollution. This study was carried out to evaluate the allelopathic potential of 92 Iranian cumin accessions grown under normal conditions and drought stress, according to the sandwich method in the laboratory. In addition, weed infestation of the same cumin accessions was examined in a field by measuring total weed species richness and diversity under normal irrigating conditions. Results indicated that length and fresh weight of radish radicle and hypocotyl are more sensitive than the other germination indices against exuded allelochemicals of cumin seeds. Furthermore, all genotypes showed the average germination percentage to be less than the control samples. They could significantly reduce total species richness and diversity in their surroundings. Weed suppressive percentage of cumin genotypes in the field experiments was positively correlated with inhibition percentage of radicle growth in the laboratory. Water stress increases the allelopathic effects of cumin accessions in laboratory screening. Classification based on principle component method indicated that cumin genotypes were categorized into two clusters in both moisture regimes. Despite having stress factors, only the genotypes 13CGOGK₁, 35CSESH₁, 65CEAAS, 68CKERF₂, 75CYAAR₂, and 84CKRTJ₂ remained in a group with strong allelopathic activity. According to these results, there is the significant diversity for allelopathic strength in the Iranian cumin germplasm, which can be useful in cumin breeding for weed suppression.  相似文献   

Interval sampling methods in Zagros forests using GIS     

Abdolali Karamshahi  Zahra Alihoseini  Javad Mirzaei  Aliakbar Jafarzadeh 《林业研究》2017,28(6):1261-1266

In this study, for accuracy and cost an optimal inventory method was examined and introduced to obtain information about Zagros forests, Iran. For this purpose,three distance sampling methods(compound, order distance and random-pairs) in 5 inventory networks(100 m × 100 m, 100 m × 150 m, 100 m × 200 m,150 m × 150 m, 200 m × 200 m) were implemented in GIS environment, and the related statistical analyses were carried out. Average tree density and canopy cover in hectare with 100% inventory were compared to each other.All the studied methods were implemented in 30 inventory points, and the implementation time of each was recorded.According to the results, the best inventory methods for estimating density and canopy cover were compound150 m × 150 m and 100 m × 100 m methods, respectively. The minimum amount of product inventory time per second(T), and(E%)2 square percent of inventory error of sampling for the compound 150 m × 150 m method regarding density in hectare was 691.8, and for the compound 100 m × 100 m method regarding canopy of 12,089 ha. It can be concluded that compound method is the best for estimating density and canopy features of the forests area.  相似文献   

Study Potential of Indigenous <Emphasis Type="Italic">Pseudomonas aeruginosa</Emphasis> and <Emphasis Type="Italic">Bacillus subtilis</Emphasis> in Bioremediation of Diesel-Contaminated Water     

Mohammad-Saeed Safdari  Hamid-Reza Kariminia  Zahra Ghobadi Nejad  Thomas H. Fletcher 《Water, air, and soil pollution》2017,228(1):37

Petroleum products which are used in a wide variety of industries as energy sources and raw materials have become a major concern in pollution of terrestrial and marine environments. The purpose of this study was to assess the potential of indigenous microbial isolates for degradation of diesel fuel. Two most proficient bacterial strains among five isolated strains from polluted soil of an industrial refinery were studied. The isolates then were identified as Pseudomonas aeruginosa and Bacillus subtilis using biochemical tests and 16S rRNA gene sequence analyses. P. aeruginosa showed higher biodegradation efficiency than B. subtilis in shaking flask containing diesel-contaminated water. P. aeruginosa and B. subtilis degraded about 87 and 75% of total hydrocarbons, respectively, in flasks containing 2% diesel and 98% water. The biodegradation efficiency of the isolates decreased as diesel contamination increased from 2 to 5%. The isolates showed significantly higher efficiency on degradation of short-chain hydrocarbons in 20 days, i.e., by using P. aeruginosa, removal efficiency of C₁₀ hydrocarbons was near 90%, while about 69% of C₂₀₊ hydrocarbons and 47% of aromatic hydrocarbons were removed. Therefore, the isolates showed high capability in biodegradation of diesel contamination of the refinery.  相似文献