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Temporal expression of three conserved putative microRNAs in response of Citrus × Limon to Xanthomonas citri subsp. citri and Xanthomonas fuscans subsp. Aurantifolii

100%
Alizadeh M., Askari H., Najafabadi M. S.
BioTechnologia. Journal of Biotechnology Computational Biology and Bionanotechnology
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2017
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tom 98
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nr 3
2

Developmental gene expression of antimicrobial peptide Protegrin-1 and effect of weaning on gene regulation of Protegrin-1 in piglets

100%
Han F. F., Wang Y. Z., Feng J., Guo J., Xu Z. R.
Journal of Animal and Feed Sciences
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2007
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tom 16
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nr 1
86-95
3

Improved fusion protein expression in EGFP via the mutation of both Kozak and the initial ATG codon

100%
Dai C., Cao Z., Wu Y., Yi H., Jiang D., Li W.
Cellular and Molecular Biology Letters
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2007
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tom 12
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nr 3
Since its discovery, green fluorescence protein (GFP) has been used as a reporter in a broad range of applications, including the determination of gene expresion in diverse organisms, and subcellular protein localization. pEGFP-N1 is a eukayotic expression vector encoding EGFP, the MCS of which locates at the N terminus of EGFP. In this study, the cDNA sequence of scorpion toxin BmKK2 was inserted into the XhoI-HindIII cut of pEGFP-N1 to construct a toxin-EGFP fusion gene (named pEGFP-BmKK2). Fluorescence imaging revealed that HEK 293T cells that were transfected by pEGFP-BmKK2 emitted green fluorescence. Transcription of pEGFP-BmKK2 was confirmed by RT-PCR. However, western blotting analysis showed that the transfected HEK 293T cells expressed mostly EGFP, but little toxin-EGFP fusion protein, implying that pEGFP-N1 cannot be used as a fusion expression vector for subcellular protein localization for the BmKK2 gene. Consequently, two modified recombinant vectors (pEGFP-BmKK2-M1 and pEGFP-BmKK2-M2) were constructed based on pEGFP-BmKK2. This greatly improved the expression of toxin-EGFP fusion protein from pEGFP-BmKK2-M2.
4

The effects of velvet antler polypeptides on the phenotype and related biological indicators of osteoarthritic rabbit chondrocytes

67%
Zhang Z., Liu X., Duan L., Li X., Zhang Y., Zhou Q.
Acta Biochimica Polonica
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2011
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tom 58
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nr 3
Objective: To study the effects of velvet antler polypeptides (VAPs) on osteoarthritic chondrocytes (OCs) in rabbits. Methods: An osteoarthritic rabbit model was established according to Hulth's method. OCs were isolated and cultured for observation of the cell cycle. Cell proliferation was detected by MTT assay and the cell cycle was monitored by flow cytometry. The phenotype was determined by toluidine blue staining as well as immunohistochemical staining for collagen type II. The expression of MMP-1, MMP-3, MMP-13, TIMP-1, and collagen I and X mRNA by chondrocytes was assayed by RT-PCR. Results: The VAPs had no obvious proliferative effect on OCs and did not affect the cell cycle. However, they significantly reduced the proportion of early apoptotic cells in a dose-dependent manner. Further, VAPs inhibited the expression of collagen I and X mRNA and induced abnormal expression of MMP-1 and MMP-13 mRNA. VAPs had no significant effect on MMP-3 and TIMP-1 mRNA levels. The toluidine blue and collagen type II immunohistochemical staining intensities of VAP-treated chondrocytes were positively correlated with the concentration of VAPs used. Conclusion: VAPs had no significant effect on OC proliferation and the cell cycle, but did increase the glycosaminoglycan (GAG) and collagen type II expression levels in the extracellular matrix, and down-regulated collagen I and X mRNA expression. Treatment of cartilage cells with VAPs maintained their normal phenotype, inhibited matrix metalloproteinases (MMPs) secretion, kept the balance of cartilage matrix metabolism, and sustained an external environment where the cartilage cells could survive. Moreover, VAPs reduced the proportion of early apoptotic cells, suggesting that they may block the apoptotic pathway in OCs.
5

Detection of jaagsiekte sheep retrovirus in respiratory tract fluid and lung tissue of experimentally infected lambs

67%
Kycko A., Jasik A., Reichert M.
Bulletin of the Veterinary Institute in Puławy
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2008
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tom 52
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nr 1
The possibility of jaagsiekte sheep retrovirus (JSRV) genome detection in peripheral blood leukocytes and respiratory tract fluid of ovine pulmonary adenocarcinoma (OPA) affected sheep was tested. Five of six lambs used in the experiment were infected with JSRV by intratracheal inoculation. The blood samples were taken from all the lambs at 10-d intervals for 3 months and the white blood cells were subjected to DNA isolation followed by PCR amplification for the detection of proviral DNA that gave negative results for all the samples. The respiratory tract fluid was collected from the nostrils of lamb No. 6, which developed clinical sign of OPA 2 months after the inoculation. The fluid was examined using PCR and reverse transcriptase PCR (RT-PCR) for the presence of proviral DNA or viral RNA. The presence of OPA in the lamb was subsequently confirmed by histopathological examination and the detection of proviral DNA in the lung tissue. Results of the standard PCR amplification performed on the DNA isolated from the nasal discharge from lamb No. 6 was negative, while the RT-PCR gave positive results confirming the presence of virions in the lung fluid. Results of the study show the RT-PCR technique may be a useful tool in ante-mortem diagnosis of OPA, especially in distinguishing it from other respiratory diseases.
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