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【Objective】 the bioinformatics software online NRD1 gene bioinformatics analysis. P19 cells are cultured in vitro and induce P19 cells to embryonic cardiomyocyte-like cells. The phenelzine lyase 1 (NRD1) p19 cells into cardiomyocyte-like cells induced differentiation process expression changes were observed. 【Methods】 using DNA Star Blast software protein physical and chemical the nature using ProtParam online tools, online the nucleotide basic characteristics analysis NRD1 gene homology; analysis; prediction of protein secondary structure to utilize online SOPMA tool to complete; Protein leader peptide and signal peptide predicted transmembrane region, hydrophobic / hydrophilic and soluble analysis of the use of online tools SignalP 3.0, TMpred ProtScale ProtParam completed; domain (domain) analysis using the SMART tool, PSORT Ⅱ predict subcellular localization; using the three-dimensional structure of the Swiss-Pdb Viewer, Gene3D, MMDB the software predicted NRD1 protein. P19 cells after recovery, cultured and passaged in vitro dimethyl sulfoxide (DMSO) induced differentiation into myocardial cells, morphological changes were observed under the light microscope, inverted microscope. p19 cells were treated with 1% dimethyl sulfoxide (DMSO) suspension induced culture 4d adherent culture to 16d, continued with growth medium, cell beating was observed by Western Blot identified cells troponin I (cTnI), and RT -PCR, Western Blot p19 cells NRD1 mRNA and protein expression. Spss 13.0 software for data management and statistical. 【Results】 NRD1 gene mRNA length 4155 bp; This gene initiation codon ATG, stop codon TAA, 29 ~ 3514 bp open reading frame; same coding frame protein translation terminator TGA before its reading frame reading frame after the polyA polyadenylation signal aataaa,; NRD1 gene is located on chromosome 4 4 ??C7 the; NRD1 containing 31 exons and 30 introns. NRD1 encoded protein of 1161 amino acids, the relative molecular mass of approximately 14,281.6 theoretical isoelectric point of 4.77, the human, mouse, rat and yeast species the gene homology; common structural elements mainly α-helix (α-helix), β-of corner (beta-turn), β-folded sheet (β-pleated sheet) and no rules crimped (random coil), etc.; α-helix and no rules a crimp is NRD1 most large number of structural elements, and the The β-turn and extended strand is spread in the whole protein; secretory pathway signal peptide (signal pep tide) score of 0.631, a signal peptide anchor score was 0.000, the maximum possible position is 19-21BP digested but score of 0.353; whole NRD1 peptide chain, there are four possible transmembrane domains; ProtScale a prediction NRD1 amino acid sequence of hydrophobic / hydrophilic, the results showed that the lowest value (-3.500) of the polypeptide chain, the highest score ( 2.667), hydrophilic amino acids are evenly distributed throughout the peptide chain and more than hydrophobic amino acids; using ProtParam http://us.expasy.org/tools/protparam html online tools to analyze NRD1 gene physical and chemical properties, and at the same time can be observed to the the gene total average hydrophilicity: -0.514, landing protein soluble analysis online server (http://biotech.ou.edu/), you can get results submitted to the amino acid sequence encoding the protein molecules results suggest NRD1 83.7% may soluble protein is soluble in water; use the SMART tools (http://smart.embl-heidelberg.de/smart/set _MODE. cgi? GENOMIC) analysis of the results showed that, NRD1 intermediate portion has three sections peptidase activity sequence, there are a lot of internal disorder sequence; use of the PSORT Ⅱ tools analysis (http://psort.nibb.ac.jp/form2.html) subcellular localization information, the results showed that 26.1% may be present in the cytoplasm, 13.0% may exist in the nucleus, 39.1% may be localized in the mitochondria, 4.3% may be located in the cytoskeleton, etc.; predicted with Swiss-Pdb Viewer, Gene3D, MMDB software NRD1 three-dimensional structure of the protein. Recovery undifferentiated P19 cells were uniform in size, scattered, small cell bodies were round, 3 ~ 4 h began adherent cells, irregular flat after 24 hours, the cells covered about 50% of the bottom of the bottle, 48h cell growth intensive. Induced by 1% dimethylsulfoxide (DMSO) 4 d, the formation of cell aggregates. Induced 4d cells adherent growth to disperse the proliferation of clonal colony. d8, you can see the pulsating cardiomyocyte-like with divergence time extension, diversification of cell morphology, increased spindle cells. p19 cells induced differentiation of the first eight days of spontaneous rhythmic beating cTnI protein positive. Gene in differentiation of NRD1 expression of 0 to 10 days was significantly higher, the difference was statistically significant (P lt; 0.05) between the point in time; 8 to 14 days of the gene expression stabilized at each time point were no differences between statistically significant (P gt; 0.05). [Conclusion] mice NRD1 gene mRNA sequences and protein sequences can be found. The acidic protein atoms mainly composed of C, H, O, N, and containing a small amount of elemental S; such as human, rat, mouse, and prokaryotes, yeast has high homology, indicating that the gene is a more important organism function relatively conserved protein; NRD1 protein does not have a leader peptide, the protein is not secreted protein may not be the signal peptide; NRD1 entire peptide chain from the four possible transmembrane domains; hydrophobicity / hydrophilicity of the analysis results may be considered NRD1 is a hydrophilic protein may be soluble proteins; three segments of the predicted internal peptidase activity sequence may make it has a peptidase activity, internal disorders sequence may make it having a folding characteristics; subcellular localization is most likely to be located in the mitochondrial, followed again cytoplasm, nucleus; presence of the protein tertiary structure is a function of protein. P19 cells induced by 1% DMSO has been given to the trend of myocardial cell differentiation, cardiomyocyte-like cells similar to embryonic formation. NRD1 gene in p19 cells expression gradually raised to myocardial cell differentiation process, may be involved in myocardial cell differentiation and development.
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