جستجوی مقالات مرتبط با کلیدواژه « بیان » در نشریات گروه « بیوتکنولوژی و ژنتیک گیاهی »

Auxin response transcription factors (ARFs) are involved in auxin-mediated responses and play a key role in regulating the growth and development of plant nutrition, such as roots, stems, leaves, and reproductive organs, such as flowers and fruits. A genome-wide analysis of ARFs can improve the understanding of their regulatory role in the growth and development of our knowledge. Although the ARF gene family has been studied in some plant species, its structural features, molecular evolution, and expression profiling in Echium plantagineum are still unknown. This study aims to better understand distinctive structural and functional features among ARF proteins in E. plantagineum.

 In this study, a comprehensive genome-wide analysis was carried out to find All members of the ARF family in E. plantagineum based on two methods 1) Hidden Markov Model (HMM) profiles of ARF gene family members and 2) Alignment with Arabidopsis (Arabidopsis thaliana) ARF genes sequence. ARF proteins were performed. The conserved motifs of ARF genes were identified, the chromosomal map and location of ARF genes were analyzed using a map chart, phylogenetic relationships using FastTree, and their protein characteristics were analyzed using the Expasy-ProtParam online server. To investigate the phylogenetic relationships of E. plantagineum ARF proteins, a phylogenetic tree was drawn using the alignment of E. plantagineum protein sequences.Functional annotation and Gene Ontology classification were determined using the g: Profiler web server. The integrated network was predicted to identify co-expressed genes using the GeneMANIA web server.

 Based on genome-wide analysis, 28 ARF genes were identified, widely distributed in multiple chromosomes of E. plantagineum. These genes have a transcription regulatory activity, and depending on the nature of the sequence, they have an activating or repressing role. Subcellular location prediction showed that the ARF proteins are most present in the nucleus. Phylogenetic analysis of the 28 ARF proteins forms ten main classes; each class is specialized in function and provides insights into different orthologous relationships. The present study identifies the ARF gene family of E. plantagineum and its evolutionary relationship with the members of this family in Arabidopsis species. This issue can help identify ARF genes and reveal their function. Analysis of conserved motifs and domain search in ARF protein sequences showed that ARF proteins have DNA binding domains such as B3 and Auxin_resp domain in their structure. Chromosomal localization analysis showed that ARF members are widely distributed in chromosomes. The analysis of gene ontology terms (GO terms) in the biological process category showed that cellular process regulation, metabolic process regulation, stimulus-response, signaling, and biological regulation are the most significant GO terms.

The results of this study provide a basis for identifying ARF genes and clarifying their function in E. plantagineum, which will be helpful for future research to discover and confirm the function of these genes.

Raini Cashmere goat is one of the most important goat breeds in Iran. These animals are breed both for meat production and cashmere production. One of the basic measures in the domestic animals is the study of genes and proteins associated with economic traits and their study at cellular or chromosomal levels. One of these important genes is the leptin. Leptin is produced by white adipose tissue and plays an important role in the regulation of feed intake, energy balance, fertility and immune functions. The aim of this research was to study leptin gene expression in adipose tissue, liver, kidney, lung and heart of Raini Cashmere goat using Real Time PCR technique.  

Tissue sampling from heart, lung, liver, kidney and adipose tissue (3 replicates from each tissue) of 6 animals was performed and RNA was extracted. Extracted RNA were immediately stored at -80°C.The Quality and quantity of RNA were evaluated and cDNA was synthesized and Real Time PCR was performed. PCR Products were electrophoresed on 1.5% agarose gel. Melting curves from Real Time PCR were examined and were evaluated different levels of expression in the studied different tissues.

The results of Real Time PCR curves and observation of electrophoresis of PCR products on agarose gel showed that the leptin gene was expressed in all the tested tissues and the highest level of expression was observed in adipose tissue (4.5) and liver (3.7) and the lowest level was detected in heart (1.4).  

These results may show that leptin plays a particular role in fat metabolism. Further studies are needed to clarify role of leptin in the physiology of fat metabolism and other materials. This would help us to better understand the mechanisms for the known effect of nutritional factors and body fatness on various functions.

The Apple chlorotic leafspot virus (ACLSV) is among of the most common virus infecting pome and stone fruit trees around the world. The virus has spread throughout the world and has been reported in many hosts such as; apples, pears, peaches and cherries. The purpose of this study is to express the coat protein (CP) of ACLSV in E. coli. CP expression in E. coli has been developed to prepare antigen for antibody production without needs to purify the virus from infected plant. In this study, complete CP gene (582 bp) from an isolate of ACLSV (SSa) was amplified by specific primers, MF and MR. The amplified CP gene was ligated to TA cloning vector (pTG19-ACLSV CP) and transformed into E. coli strain DH5α. Transformed cells were selected on LB containing ampicillin, X-Gal and IPTG, and recombinant plasmids were confirmed by restriction analysis. Then, the pTG-ACLSV CP was sub-cloned into pET28a (+) as expression vector that digested by BamHl and XhoI restriction enzymes. Finally, the pET28a-ACLSV CP was transformed into E. coli strain BL21 by heat shock method. For ACLSV CP expression, the cells containing pET28a-ACLSV CP were induced by 1 mM of IPTG and the protein was extracted two and four hours after induction and analyzed in SDS-PAGE. The SDS-PAGE result showed that ACLSV CP have been expressed that is expected based on additional tags from plasmid.

 In dairy cattle, the increase in milk yield has been accompanied by a more negative energy balance during early lactation and a decrease in fertility. As the hormone leptin is involved in regulation of nutritional status and reproductive function, this hormone is an interesting protein to investigate during the periparturient period in dairy cattle when many changes take place both in energy metabolism and reproductive physiology. The aim of this research was to study leptin gene expression in adipose tissue of Holstein dairy cattle.   Tissue sampling from subcutaneous adipose tissue of 20 Holstein cattle on the 20th day of lactation, with the same gestational age (second pregnancy) and mean body weight of 680 ± 80 kg was performed and RNA was extracted. Extracted RNA were immediately stored at -80°C.The Quality and quantity of RNA were evaluated and cDNA was synthesized and Real Time PCR was performed. PCR Products were electrophoresed on 2% agarose gel and were evaluated level of gene expression in the studied tissue.   Results showed that the leptin gene was expressed in the subcutaneous adipose tissue. These results may show that leptin plays a particular role in fat metabolism.   During the periparturient period the cow mobilizes her fat reserves (i.e. adipose tissue) and it appears that all her energy is going to the production of milk, and that other processes like reproduction and immunity, get a lower priority. Because fertility, just as milk production, is also an economically important trait, fertility should be considered as part of a dairy cattle breeding program. Generally, further studies are needed to clarify role of leptin in the physiology of fat metabolism and other materials. This would help us to better understand the mechanisms for the known effect of nutritional factors and body fatness on various functions.

Leptin is produced by white adipose tissue and plays an important role in regulation of feed intake, energy balance, fertility and immune functions. The aim of this research was to study leptin gene expression in adipose tissue, liver, kidney, lung and heart of Kermani sheep. Tissue samples obtained from 6 animals and RNA was extracted. Extracted RNA were immediately stored at -80°C. Quality and quantity of RNA were evaluated and cDNA was synthesized and Real Time PCR was performed. PCR Products were electrophoresed on 1.5% agarose gel and were evaluated different levels of expression in studied different tissues. Results showed that the leptin gene was expressed in all the tested tissues and the highest level of expression was observed in adipose tissue and liver and the lowest level was detected in heart. These results may show that leptin plays a particular role in fat metabolism. Further studies are needed to clarify role of leptin in the physiology of fat metabolism and other materials. This would help us to better understand the mechanisms for the known effect of nutritional factors and body fatness on various functions.

Because of limitations in antibody production against plant viruses, coat protein expression has been developed to prepare antigen for antibody production. In the recent research, Nucleocapsid gene of Tomato spotted wilt virus was amplified from a local strain in Iran. Cloned segment in TA vector (pTG19TSWV-N) was sub-cloned into pET32a as expression vector that digested by XhoI and BamHI. Then, pET32TSWV-N was transformed in two different strains of E. coli BL21(DE3) and BL21(DE3) pLysS by heat shock method. For protein expression, the transformed cells were induced by 1mM of IPTG in both strains. The protein was extracted four hours after induction and analyzed in SDS-PAGE. The SDS-PAGE result showed that a 48 kDa protein have been expressed that is expected based on additional tags from plasmid. The result revealed that nucleocapsid had been expressed in both strains whereas protein expression was little more in BL21(DE3) pLysS.

Drought stress is one of the most important limiting factors of crop plants in Iran and worldwide. To assess the response of mutant line MT149 to drought stress in comparison to parental cultivar Neda, a factorial experiment was conducted in a completely randomized design. Drought stress (-6 bar) was induced using poly ethylene glycol 6000 and leaf growth dynamics and chlorophyll a and b contents were evaluated in different time series (0, 12, 24 and 36 hours after stress treatment). Results showed that effects of genotype, time and stress were significant on all investigated characters. Also, mean`s comparisons showed that mutant line MT149 was superior over than itself parental counterpart. Furthermore, assessment of catalase gene isoform A (OsCat A) in different times after stress showed that parental cultivar Neda experienced a significant down-regulation of catalase A gene, while MT149 experienced a significant up-regulation of the gene in the same time. Based on the results of this research it can be concluded that one of evidences on a higher drought tolerance of MT149 is probably up-regulation of OsCat A to scavenge the ROS toxicities.

Rheb (Ras homolog enriched in brain) gene is originally identified as immediate early gene (IEG) in 1994, encoding 184 amino acids with a deduced molecular mass of 20 497 Da in hippocampus. Rheb belongs to Ras family that encodes a carboxylterminal CAAX box indicating that the protein may undergo post-translational farnesylation. Rheb is an upstream regulatory factor in mTOR signaling pathway and the Rheb-GTP can active mTOR that inducing strong phosphorylation of endogenous S6K1 at residues Thr389, Thr421 and Ser424. Overexpression of Rheb stimulates cell growth while knockdown of Rheb expression inhibits protein synthesis and cell growth in insects. Over expression of Rheb-GAP inhibits mTOR activation and reduces fiber cross-sectional area in mammalian skeletal muscle. There is cross-talk between Mstn and mTOR signaling pathways in mammalian skeletal muscles. Tissues including brain, heart, lung, pancreatic, spleen, kidney, liver and testis were collected from the Raini Cashmir goat after slaughter. Extracted RNA were immediately stored at - 80°C. Quality and quantity of RNA were evaluated and cDNA was synthesized and PCR was performed. PCR Products were electrophoresed on 1.5% agarose gel and were evaluated different levels of expression in studied different tissues. Results showed that the Rheb gene was expressed in all the tested tissues and the highest level of expression was observed in kidney and the lowest level was detected in pancreatic and testis. Results of SPSS analyziz demonstrated that expression of Rheb gene in Raini cashmir goat significantly (P£0.01) is different in various tissues. Hence, can suggest that Rheb has probably role in goat cells and must detect in future investigations.