Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UNIPROT:P02794 (ferritin)
 17,525 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Our previous work showed that immunization of mice with Schistosoma japonicum (Sj) immature eggs induced significant immunity against fecundity and embryonation of the parasite. The Sj adult cDNA library was screened by sera from rabbits against Sj immature egg antigen (RASjIEA). The genes encoding molecules which may induce immunity against fecundity/embryonation were chosen for further cloning and expression. First of all, RASjIEA was absorbed with E. coli lysate to remove cross reactive antibodies. The cDNA library was then immunoscreened using the routine method. The resulted positive plaques were rescreened till individual clones were confirmed. Phagemids were obtained using in vivo excision. The positive clones were amplified using PCR. The sizes of the genes were determined by agarose gel electrophoresis. After DNA sequencing of the genes cloned, Gene bank was searched and six different genes were identified from a total of 102 positive clones. One of six identified genes, Sj ferritin (SjFer) was chosen to subclone into pGMC vector. According to DNA sequences of Sj Fer and MCS (multiple cloning site) of the vector, forward primer (Fer/GMC1) and reverse primer (Fer/GMC2) were designed and used to amplify Sj Fer by PCR. The Sj Fer cDNA and expression vector pGMC were digested with BamHI and XhoI. The digested cDNA and pGMC were ligased by T4 DNA ligase to construct a recombinant which was then used to transform E. coli strain ER2566. The fusion protein GMCSF-Sj Ferritin was expressed in insoluble form, the inclusion body. Pellets were harvested and resolved in Tris-HCl buffer containing 8M urea. GMCSF-Sj Ferritin was purified by affinity chromatography using Ni-NTA resin. The molecular weight was determined by SDS-PAGE. This study first reports the gene encoding S. japonicum ferritin as a new candidate for schistosome vaccine.
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PMID:[Schistosoma japonicum ferritin: cloning, nucleotide sequencing, expression, and purification]. 1068 50

We have investigated the analysis of RNA by use of terminal transferase-dependent PCR (TDPCR), a procedure previously used for the analysis of DNA and chromatin [J. Komura and A.D.Riggs, Nucleic Acids Res.,26, 1807-1811 (1998)]. When preceded by reverse transcription (RT), TDPCR provides an extremely sensitive, versatile, quantitative and nucleotide-level assay for detecting RNA lesions or structures that block primer extension during the RT step. The procedure is: (i) RT using a gene-specific oligonucleotide; (ii) ribo-tailing of the single-stranded cDNA product by use of terminal deoxy-nucleotidyl transferase; (iii) ligation of a DNA linker to the tailed cDNA by use of T4 DNA ligase; and (iv) PCR using a nested, gene-specific primer and a linker-specific primer. This procedure combines the versatility of a primer extension assay with nucleotide-level resolution, the specificity of nested primers and the sensitivity of PCR. Band patterns obtained are reproducible and quantifiable. We successfully used the technique for the study of yeast RNA structure, splicing intermediates and ribozyme cleavage. Also, in vivo footprint experiments, using mammalian cells and RNase T1, revealed the binding of iron-responsive element binding protein to iron responsive elements in the mRNAs of transferrin receptor and ferritin H-chain.
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PMID:In vivo, high-resolution analysis of yeast and mammalian RNA-protein interactions, RNA structure, RNA splicing and ribozyme cleavage by use of terminal transferase-dependent PCR. 1071 Apr 33