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Cell and Molecular Research - Volume:9 Issue: 1, Summer and Autumn 2017

Journal of Cell and Molecular Research
Volume:9 Issue: 1, Summer and Autumn 2017

  • تاریخ انتشار: 1396/02/22
  • تعداد عناوین: 7
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  • Jafar Vatandoost, Shadi Tirdad Pages 1-5
    In the production of recombinant proteins, the selection of an expression system is very important. Although CHO cells are used as mammalian expression system, the ability of insect Schneider line 2 (S2) cells as a new expression system for the high production of many human proteins were confirmed. It is suggested that high copy number of an introduced gene and so high transcription in these cells can be regarded as one of the reasons for high expression of recombinant proteins. Therefore, the present study aimed to evaluate the correlation of recombinant human coagulation factor IX (hFIX) abundance and mRNA expression. The amount of hFIX mRNA by quantitative real-time PCR as well as hFIX protein in cell lysate and cultured media by Elisa was analyzed. The results of data analysis indicated 6 fold increases in mRNA level in S2 cells in comparison to CHO cells. Furthermore, S2 cell line indicated 5.5 and 7-fold increase in total and secreted protein level, respectively, compared to CHO cell line. The data demonstrated the correlation of mRNA and protein abundance and indicate that S2 cell lines are superior in producing the recombinant proteins.
    Keywords: Drosophila S2 cells, CHO cells, Coagulation factor IX, Real-time PCR, Elisa
  • Soheila Shokrollahzade, Arshad Hosseini, Majid Safa, Shamim Sarhadi Pages 6-15
    MicroRNAs by their structural complementarity capabilities have canonical roles in gene regulation. In this paper; we investigate expression of EGFR, MAP2K4 and E2F3 genes targeted by miR-141, a member of miR-200 family. EGFR, MAP2K4 and E2F3 were predicted as the potential targets of mir-141 by using online miRNA bioinformatics tools. MCF-7 cells were transfected with mir-141-precursor and inhibitor vectors. Expression of miR-141 and target genes was determined by using qRT-PCR. To see the most relevant pathways regulated by miR-141, we constructed two separate networks by NetworkAnalyst and enriched list of underlying genes by Enrichment analysis tools. The expression changes of all three predicted targets were higher in transfected cells with anti-mir-141 vector, compared with the control untransfected cells. By contrast, in transfected cells with pre-mir 141, we did not see significant expression changes in EGFR, E2F3 and MAP2K4. List of genes in total networks as well as explored functional modules were enriched separately. Enrichment analysis shows that immune system pathway has the strongest relationship with the proteins potentially targeted by miR-141. The present study demonstrated potential role of miR-141 in regulation of EGFR, MAP2K4 and E2F3 expression and suggested innate immunity pathways as the key pathway through which this regulatory network contributes to breast cancer development.
    Keywords: Breast cancer, miR-141, EGFR, MAP2K4, E2F3
  • Ayeh Sadat Sadr, Mohammadreza Nassiri, Seyed Alireza Salami, Mohammad Reza Bakhtiarizadeh, Mojtaba Tahmoorespur, Alireza Shafeinia Pages 16-22
    The poultry industry occupies an important position in the provision of animal protein. Recently, next generation sequencing technology (RNA-Seq) has become available as a powerful tool to investigate transcriptional profiles for gene expression analysis of many organisms. The main use of RNA-Seq in agriculture species are focusing on finding the immune related genes or pathways by comparison of the whole transcriptome following pathogen challenge. Alternative splicing (AS) is the major fundamental mechanism generating the protein diversity and regulating the gene expression in eukaryotic organism. Identifying genes that are differentially spliced between two groups of RNA-sequencing samples is interesting subject in transcriptome with next-generation sequencing technology in this study used RNA sequencing to comparison isoforms of two breeds. A total of 64,819 transcripts were identified by aligning sequence reads to genome among the evaluated isoforms for expression analysis, 310 were significantly differentially expressed between two breeds, including 251 up-regulated and 59 down-regulated. The KEGG results of up regulated isoforms showed that that no pathway was found significantly different (FDR ≤ 0.05). However, enrichment analysis suggested that seven were over-represented (P-value ≤ 0.05) within the up regulated isoforms. Only one of them functionally related to immune system, natural killer cell mediated cytotoxicity. The results showed genes which are breed-specific expression and the comparative transcriptome analysis help to understand the difference of genetic mechanism.
    Keywords: Isoforms, RNASeq, Poultry
  • Behnaz Safamanesh, Sedigheh Esmaeilzadeh Bahabadi, Ali Izanloo Pages 23-34
    Barberry fruit is a medicinal plant, and it is one of the most important horticultural crops in South Khorasan province, Iran. Genetic diversity has a basic role in the successful breeding of crop varieties with durable resistance to biotic and abiotic stresses. The main objective of this study was to assess the genetic diversity of 20 ecotypes of Berberis, which collected from different regions of South Khorasan province, Iran, using ISSR and SSR markers. In this study, 10 ISSR primers and 5 SSR primer pairs, with the amplification of suitable polymorphic alleles were used. A total of 98 bands for ISSR markers and 43 bands for SSR markers were detected between 300 to 1300 bp and 100 to 1100 bp in size, respectively. Polymorphic ISSR-7 and CA03 primers amplified the highest number of alleles with 17 and 24 bands, respectively, while ISCS50 and CA30 primers amplified only two polymorphic alleles. The ISCS57 and GA31 primers had the highest polymorphic information content (PIC) and ISCS50 and GA04 primers had the lowest PIC. The estimated Nei's and Shannon indices for genetic diversity in ISSR markers were 0.24 and 0.35, while for SSR markers these were 0.23 and 0.34, respectively. Based on cluster analysis, five and six main groups were identified for ISSR and SSR markers, showing high genetic variations among a set of collected barberry ecotypes. Analysis of molecular variances in both ISSR and SSR markers showed that high level of total variation was due to within populations, rather. Therefore, it will be better to select within populations in breeding programs.
    Keywords: Berberis spp, Genetic diversity, ISSR, SSR
  • Mahsa Alem, Reza Asadpour, Raziallah Jafari Joozani, Katayoon Nofouzi Pages 35-38
    Enzootic abortion of ewes (EAE) induced by Chlamydophila abortus (formerly Chlamydia psittaci serotype1) is a major cause of reproductive failures in most sheep producing countries. In this study, the abortion prevalence of Chlamydophila abortus (C. abortus) in sheep abortion cases are evaluated by polymerase chain reaction (PCR) and objectives are considered Chlamydiosis incidence of abortion in sheep in the northwest region of Iran. For this purpose, the contents of fetal tissue from 50 aborted fetuses were homogenized and DNA extracted from them and then PCR is done using specific primers for Chlamydophila (CHOMP191, CHOMP371). The study showed that about 26% of the total sample is contained Chlamydophila. This study confirms that C. abortus as an important cause of ovine abortion in the northwest of Iran and showed the PCR in tissue pools of aborted fetuses is a useful method for rapid detection of Chlamydophila abortus Ovis infections.
    Keywords: Chlamydophila abortus, Molecular detection, Polymerase Chain Reaction (PCR)
  • Ehsan Nasr, Hrachya Hovhannisyan, Pourkazemi Mohammad Pages 39-43
    The biological activity of Persian sturgeon (PS) enfolded crude recombinant growth hormone (rGH) recovered from inclusion bodies (IB) estimated by administrating to fish fingerlings. The psrGH IBs expressed in E. coli were dissolved in cold guanidine HCl solution, immediately diluted ~1000 fold in cold sodium chloride 0.9% solution do not allow the rGH folding and administrated to fishes. It was revealed that intramuscularly injections of rGH at dosage of 0.5 μg/ g once a week for 8 weeks significantly accelerate the growth of fishes. Comparison of the mean growth rate and daily weight and length gain dates of Persian sturgeon fingerlings administrated by pure and crude grade rGH revealed the existence at list 2-5% potentially biologically active psrGH molecules in IBs. This fact enables to avoid the time and cost consuming processes of refolding and purification of rGHs.
    Keywords: Inclusion body, Persian sturgeon, Recombinant growth hormone, Biological activity
  • Atieh Teymoori, Mojtaba Teimoori, Madjid Momeni-Moghaddam Pages 44-53
    For 50 years, the term gene is synonymous with regions of the genome gene that coding by mRNAs and translate to protein. nonetheless, Genome wide Recent studies have revealed that regulating gene expression through degradation or translational inhibition of their point mRNAs and thus attend in a wide variety of physiological and pathological cellular processes including: development, cell proliferation, differentiation, and apoptosis pathways by thousands of regulatory non coding RNA such as lncRNAs and microRNAs. According to a recent survey, it is known this RNAs have vital role in regulation cellular pathways at transcriptional, posttranscriptional and epigenetic levels. These noncoding genes are often aberrantly expressed in a variety of human cancers. However, the biological functions of most ncRNAs remain largely in doubt. In this review, we proved that a remarkable part of the genetic etiology of cancer is imposed by noncoding regulatory sequences. The purpose of this review is aimed to give an outlook of using of noncoding RNA as diagnostic markers and therapeutic targets. These observations emphasized that the recognition of coding genes and Research continued evolution and function of non-coding RNAs for a comprehensive understanding human complex diseases like cancer are essential.
    Keywords: NcRNA, Expression, Transcription, Cell proliferation, Apoptosis, Cancer