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        Genome-wide identification and comparative analysis of MATE gene family in Cucurbitaceae species and their regulatory role in melon (Cucumis melo) under salt stress

        Shah Iftikhar Hussain,Manzoor Muhammad Aamir,Sabir Irfan Ali,Ashraf Muhammad,Haq Fazal,Arif Samiah,Abdullah Muhammad,Niu Qingliang,Zhang Yidong 한국원예학회 2022 Horticulture, Environment, and Biotechnology Vol.63 No.4

        The multidrug and toxic compound extrusion (MATE) protein family includes a primeval gene family of secondary transporters that export toxins, extrude metabolites, and participate in plant defense mechanisms. However, lack of information regarding the MATE gene family in Cucurbitaceae, a comprehensive genome-wide analysis of the MATE family was carried out in four Cucurbitaceae species (Cucumis melo, Cucumis sativus, Cucurbita pepo, and Lagenaria siceraria), and 174 MATE genes were identified. Phylogenetic and structural analysis revealed that the Cucurbitaceae MATE transporters family could be further classified into seven subgroups (A–G). GO annotation-based subcellular localization analysis predicted that most of the MATE gene family members localized on the plasma membrane. Moreover, conserved motifs and gene structure (intron/exon) analysis revealed the functional divergence between clades. Transposed duplication events have played a key role in the expansion and evolution of the MATE gene family in Cucumis melo. Cis-acting elements analysis of MATE family genes revealed that these could be targeted by a diverse set of trans-acting factors involving the MATE gene family to manage diverse stress conditions. The Chromosomal localization and molecular characteristics (weight, length, and pI) were performed using numerous bioinformatics tools. Intraspecies microsynteny analysis demonstrated that maximum orthologous genes were found between A. thaliana, C. pepo, C. lanatus, and L. siceraria. Further, functional analysis of microRNAs demonstrated miRNAs are involved in the growth and regulation of MATE genes. Finally, eleven candidates MATE genes were selected randomly, and their expression analysis was carried out via qRT-PCR at 0 h and after 24 h of salt stress. Furthermore, transient CmMATE expression in Arabidopsis thaliana protoplasts showed that protein localized on the plasma membrane. This study provides insights into the functional analysis of the MATE gene family in Cucurbitaceae species and laid down the basic knowledge to explore the role and mechanism of the MATE gene family to cope with severe salt stress conditions.

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        Aberrant Promoter Methylation at CpG Cytosines Induce the Upregulation of the E2F5 Gene in Breast Cancer

        Arshad Ali,Farman Ullah,Irum Sabir Ali,Ahmad Faraz,Mumtaz Khan,Syed Tahir Ali Shah,Nawab Ali,Muhammad Saeed 한국유방암학회 2016 Journal of breast cancer Vol.19 No.2

        Purpose: The promoter methylation status of cell cycle regulatory genes plays a crucial role in the regulation of the eukaryotic cell cycle. CpG cytosines are actively subjected to methylation during tumorigenesis, resulting in gain/loss of function. E2F5 gene has growth repressive activities; various studies suggest its involvement in tumorigenesis. This study aims to investigate the epigenetic regulation of E2F5 in breast cancer to better understand tumor biology. Methods: The promoter methylation status of 50 breast tumor tissues and adjacent normal control tissues was analyzed. mRNA expression was determined using SYBR® green quantitative polymerase chain reaction (PCR), and methylation- specific PCR was performed for bisulfite-modified genomic DNA using E2F5-specific primers to assess promoter methylation. Data was statistically analyzed. Results: Significant (p<0.001) upregulation was observed in E2F5 expression among tumor tissues, relative to the control group. These samples were hypo-methylated at the E2F5 promoter region in the tumor tissues, compared to the control. Change in the methylation status (Δmeth) was significantly lower (p=0.022) in the tumor samples, indicating possible involvement in tumorigenesis. Patients at the postmenopausal stage showed higher methylation (75%) than those at the premenopausal stage (23.1%). Interestingly, methylation levels gradually increased from the early to the advanced stages of the disease (p<0.001), which suggests a putative role of E2F5 methylation in disease progression that can significantly modulate tumor biology at more advanced stage and at postmenopausal age (Pearson’s r=0.99 and 0.86, respectively). Among tissues with different histological status, methylation frequency was higher in invasive lobular carcinoma (80.0%), followed by invasive ductal carcinoma (46.7%) and ductal carcinoma in situ (20.0%). Conclusion: Methylation is an important epigenetic factor that might be involved in the upregulation of E2F5 gene in tumor tissues, which can be used as a prognostic marker for breast cancer.

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