ABSTRACT
MicroRNAs (miRNAs) act as regulators of gene expression by binding to the 3’ untranslated region (UTR) of target genes. They perform important biological functions in the various species. Among many miRNAs, miR-21-3p is known to serve vital functions in development and apoptosis in olive flounder. Using genomic and bioinformatic tools, evolutionary conservation of miR-21-3p was examined in various species, and expression pattern was analyzed in olive flounder. Conserved sequences (5’-CAGUCG-3’) in numerous species were detected through the stem-loop structure of miR-21-3p. Thus, we analyzed target genes of miR-21-3p. Among them, 3’ UTR region of PPIL2 gene indicated the highest binding affinity with miR-21-3p based on the minimum free energy value. The PPIL2 gene showed high expression levels in testis tissue of the olive flounder, whereas miR-21-3p showed rather ubiquitous expression patterns except in testis tissue, indicating that miR-21-3p seems to control the PPIL2 gene expression in a complementary repression manner in various tissues of olive flounder. Taken together, this current study contributes to infer the target gene candidates for the miR-21-3p using bioinformatics tools. Furthermore, our data offers important information on the relationship between miR-21-3p and target gene for further functional study.
Subject(s)
Apoptosis , Computational Biology , Conserved Sequence , Flounder , Gene Expression , MicroRNAs , Olea , Repression, Psychology , Testis , Untranslated RegionsABSTRACT
Background Epidemic keratoconjunctivitis is a common eye disease,and adenovirus is one of the common pathogens.The hexon protein,one main capsid protein of the virus,is an important target of antibody binding.Thus,sequencing the coding region of the hexon protein is an important way for adenovirus fast typing.Objective This study was to complete a molecular epidemiology survey of epidemic keratoconjunctivitis and investigate its association with adenovirus in Shanghai area by sequencing the coding region of hexon protein.Methods Two hundred and fourteen sacconjunctival swab specimens were collected from 214 patients with suspicious epidemic keratoconjunctivitis who visited Shanghai Eye Disease Prevention and Treatment Center and the clinical sites supervised by the Shanghai Prevention and Monitoring Office of Acute Hemerragic Conjunctivitis under the informed consent from January 2010 to December 2012.DNA was extracted from the specimens and then the 140 bp conserved sequence in hexon protein coding region was amplified by PCR initially to determine an adenovirus pathogen.Furtherly,956 bp conserved sequence of the hexon codind district was sequencied to clarify the serotype of adenovirus in the adenovirus-positive specimens.Results 50.93％ patients (109/214) were detected to be adenovirus-positive by generic PCR,in which AdV1 + was in 4 patiens,AdV2+ was in 33 patients,AdV3+ was in 15 patients,AdV4+ was in 12 patients,AdV8+ was in 19 patiens,AdV19+ was in 15 patients,AdV37+ was in 8 patients.The subgenus D adenoviruses,including AdV8+,AdV19+ and AdV37+ often resulted in corneal inflammation,pseudomembranous conjunctivitis and preauricular lymph nodes;while subgenus B adenovirus induced much frequent tract infection and less corneal response.Conclusions PCR-sequence of conserved region of hexon protein coding district is applicable for the detection and serotyping of adenovirus in epidemic keratoconjunctivitis.
ABSTRACT
Objective To investigate and detect the immunity characteristics of a conserved sequence including 30 amino residues which is located in the C terminal of HPVL1 .Methods The immune model of mouse was establish with a polypeptide synthetized based on this sequence .The amount of CD3+ ,CD3+CD4+ or CD3+CD8+ lymphocyte of the mouse spleen was detected by Flow cy-tometry ,then the value of CD4+ /CD8+ were calculated .The cell proliferation was detected by MTT assay .Sample was took from the cell supernatant to detect the content of IL-4 and IFN-γby double antibody sandwich ELISA .Results (1)The amount of CD3+CD8+ lymphocytes and the value of CD4+ /CD8+ were significantly increased than control group(P0 .05) .(3)The level of IL 4 were significantly higher than that of control group (P0 .05) .Conclusion The results showed that it was affirmative that the polypeptides induced humoral immunity ,which was worth to be furtherly studied as a preventive vaccine .But its cell immunogenicity is weak ,and the attenmpt to induce the response related to the cell immunogenicity was failed ,and the immunogenicity of the peptide remains to be improved .
ABSTRACT
190-kilodalton glycoprotein (P190) of Plasmodium falciparum. precursor of the major surface protein of merozoites, is considered a promising candidate for blood stage malarial vaccine. We designed six primers according to the sequence of MAD20 strain, with a GC clamp and BamHI site at the 5'- end of each one, and a GC clamp and Xbal site at the 3'- end of each one. The primers were synthesized by phosphoramidite approach (User's Manual of ABI Company) and purified using HPLC. Three fragments in the second, third and fourth conserved regions of P190 gene of Plasmodium falciparum FCC1/HN strain isolated from the blood of patients in Hainan Province of China were amplified by the polymerase chain reaction (PCR) technique. The amplified fragments were subcloned into pUC18 vectors and sequenced using the dideoxy chain termination method. All three regions of P190 gene of FCCl/HN strain also were highly conservative as compared with P190 gene of MAD20 (Papua New Guinea isolate), K1 (Thailand isolate), Wellcome (West Africa isolate) and CAMP (Malaysia) strains of Plasmodium falciparum. The C at position 81 in the second conserved block of P190 gene of FCC1/HN isolate was substituted by T, which did not change the amino acid determined by the coden corresponding to the substitution.