OBJECTIVETo type and group the Yersinia pestis strains isolated in China to clarify the geographical distribution of ribotypes of Yersinia pestis.

METHODSGenomic DNA of Yersinia pestis were digested with EcoR I, then hybridized with 16s-23s-5s rRNA gene probe.

RESULTSThese tested strains were divided into 3 ribotypes, the profiles obtained were relatively homogeneous, with most of them differed only by the presence or the absence of 1 - 2 restriction fragments. Ribotype A and B were the most common types, which distributed in a large area in China while ribotype C was the least, only limited to a small area. There was certain correlation between the ribotypes and the plague foci, usually only one ribotype was found in one plague foci.

CONCLUSIONThe ribotypes were stable in the plague foci. Correlation between the ribotypes of Yersinia pestis strains and their geographical origins was noticed. All 3 ribotypes had different origins, however ribotype A and ribotype C seemed to be closer related.

China ; DNA, Bacterial ; genetics ; Geography ; RNA, Ribosomal, 16S ; genetics ; RNA, Ribosomal, 23S ; genetics ; RNA, Ribosomal, 5S ; genetics ; Ribotyping ; Yersinia pestis ; classification ; genetics

Objective To describe the microbiological characteristics of ()CGMCC 12426 and determine and analyze its complete genome sequences.Methods strain CGMCC 12426 genomic DNA sequencing was performed on a single molecule real-time sequencing(SMRT)platform and the annotation was completed in the NCBI Prokaryotic Genomic Annotation Pipeline(pGAP).Results The complete genomic sequences of the released CGMCC 12426 consisted of a 4 138 265-bp circular chromosome and a 74 165-bp plasmid,which resulted in the prediction of 4581 genes including 4222 coding sequences,87 tRNAs,and 30 rRNAs(which included 5S rRNA,16S rRNA,and 23S rRNA).Conclusion The genome sequencing provided a basis for further investigations on the genetic background of and on the metabolic and regulatory mechanisms.
Bacillus subtilis ; genetics ; Genome, Bacterial ; Plasmids ; RNA, Ribosomal, 16S ; genetics ; RNA, Ribosomal, 23S ; genetics ; RNA, Ribosomal, 5S ; genetics ; Sequence Analysis, DNA

OBJECTIVEThis research focused on analyzing the differences of 5S rRNA gene spacer sequences on Swertia mussotii and its commonly used adulterants, including S. franchetiana, S. wolfangiana and S. chirayita.

METHODDNA was extracted from the collected Swertia samples. 5S rRNA intergenic spacers were amplified by PCR, sequenced and analyzed.

RESULT5S rRNA gene spacer sequences were different between S. mussotii and its other three adulterants. Sequence divergence among species ranged from 30.6% to 65.0%.

CONCLUSION5S rRNA spacers may be used as molecular authentication markers to differentiate S. mussotii and other commonly used Swertia adulterants. This result provides reliable and simple reference for the authentication of Swertia genus species.

Base Sequence ; Molecular Sequence Data ; Polymerase Chain Reaction ; RNA, Ribosomal, 5S ; genetics ; Sequence Homology, Nucleic Acid ; Swertia ; classification ; genetics

DNA, Ribosomal ; genetics ; Genome, Plant ; Phylogeny ; Polymerase Chain Reaction ; methods ; Polymorphism, Single Nucleotide ; genetics ; RNA, Ribosomal, 5S ; genetics ; Sequence Homology, Nucleic Acid ; Species Specificity ; Valerian ; classification ; genetics

OBJECTIVES: To explore the correlation between the expression levels of several RNA markers in human brain tissue and early postmortem interval （PMI）. METHODS: Twelve individuals with known PMI （range from 4.3 to 22.5 h） were selected and total RNA was extracted from brain tissue. Eight commonly used RNA markers were chosen including β-actin, GAPDH, RPS29, 18S rRNA, 5S rRNA, U6 snRNA, miRNA-9 and miRNA-125b, and the expression levels were detected in brain tissue by real-time fluorescent quantitative PCR. The internal reference markers with stable expression in early PMI were screened using geNorm software and the relationship between its expression level and some relevant factors such as age, gender and cause of death were analyzed. RNA markers normalized by internal reference were inserted into the mathematic model established by previous research for PMI estimation using R software. Model quality was judged by the error rate calculated with estimated PMI. RESULTS: 5S rRNA, miRNA-9 and miRNA-125b showed quite stable expression and their expression levels had no relation with age, gender and cause of death. The error rate of estimated PMI using β-actin was 24.6%, while GAPDH was 41.0%. CONCLUSIONS 5S rRNA, miRNA-9 and miRNA-125b are suitable as internal reference markers of human brain tissue owing to their stable expression in early PMI. The expression level of β-actin correlates well with PMI, which can be used as an additional index for early PMI estimation.
Actins/analysis* ; Autopsy ; Brain/metabolism* ; Humans ; MicroRNAs/analysis* ; Models, Theoretical ; Postmortem Changes ; RNA Stability ; RNA, Ribosomal, 18S/analysis* ; RNA, Ribosomal, 5S/analysis* ; RNA, Small Nuclear/analysis* ; Real-Time Polymerase Chain Reaction ; Software

OBJECTIVETo define the main genotypes in Guizhou agricultural areas by molecular epidemiologic investigation of 21 Borrelia burgdorferi sensu lato of Lyme disease spirochetes and to provide the scientific bases for formulating a preventive policy.

METHODSPolymerase chain reaction (PCR) technique was used to amplify the 23S(rrl)-5S(rrf) intergenic spacer, and amplified products were analyzed by restriction fragment length polymorphism (RFLP) and nucleotide sequencing.

RESULTSThere were two genospecies in the strains: 20 strains belong to Borrelia valaisiana, 1 strain is Borelia sp.

CONCLUSIONBorrelia valaisiana was the main genotype in Guizhou agricultural areas. The harmness of B. valaisiana to human being has been confirmed. In order to efficiently prevent the harmness of agent to the people in Guizhou agriculture areas, we should study the risk further.

Base Sequence ; Borrelia burgdorferi ; classification ; genetics ; China ; DNA, Bacterial ; chemistry ; genetics ; DNA, Ribosomal Spacer ; genetics ; Humans ; Lyme Disease ; microbiology ; Molecular Sequence Data ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; RNA, Ribosomal, 23S ; genetics ; RNA, Ribosomal, 5S ; genetics ; Ribotyping ; Sequence Analysis, DNA

The first human case with trichinellosis was reported in 1964 in Tibet, China. However, up to the present, the etiological agent of trichinellosis has been unclear. The aim of this study was to identify a Tibet Trichinella isolate at a species level by PCR-based methods. Multiplex PCR revealed amplicon of the expected size (173 bp) for Trichinella spiralis in assays containing larval DNA from Tibet Trichinella isolate from a naturally infected pig. The Tibet Trichinella isolate was also identified by PCR amplification of the 5S ribosomal DNA intergenic spacer region (5S ISR) and mitochondrial large-subunit ribosomal RNA (mt-lsrDNA) gene sequences. The results showed that 2 DNA fragments (749 bp and 445 bp) of the Tibet Trichinella isolate were identical to that of the reference isolates of T. spiralis. The Tibet Trichinella isolate might be classifiable to T. spiralis. This is the first report on T. spiralis in southwestern China.
Animals ; DNA, Helminth/chemistry/genetics ; DNA, Mitochondrial/chemistry/genetics ; DNA, Ribosomal/chemistry/genetics ; DNA, Ribosomal Spacer/genetics ; Genotype ; Humans ; Multiplex Polymerase Chain Reaction ; RNA, Ribosomal, 5S/genetics ; Sequence Analysis, DNA ; Swine ; Swine Diseases/*parasitology ; Tibet ; Trichinella spiralis/*classification/genetics/isolation & purification ; Trichinellosis/parasitology/*veterinary

A simple, rapid, and high-throughput method for detection and identification of Wuchereria bancrofti, Brugia malayi, Brugia pahangi, and Dirofilaria immitis in mosquito vectors and blood samples was developed using a real-time PCR combined with high-resolution melting (HRM) analysis. Amplicons of the 4 filarial species were generated from 5S rRNA and spliced leader sequences by the real-time PCR and their melting temperatures were determined by the HRM method. Melting of amplicons from W. bancrofti, B. malayi, D. immitis, and B. pahangi peaked at 81.5+/-0.2degrees C, 79.0+/-0.3degrees C, 76.8+/-0.1degrees C, and 79.9+/-0.1degrees C, respectively. This assay is relatively cheap since it does not require synthesis of hybridization probes. Its sensitivity and specificity were 100%. It is a rapid and technically simple approach, and an important tool for population surveys as well as molecular xenomonitoring of parasites in vectors.
Animals ; Blood/*parasitology ; Brugia/classification/genetics/*isolation & purification ; Cats ; Culicidae/*parasitology ; Dirofilaria immitis/classification/genetics/*isolation & purification ; Dogs ; Humans ; Male ; Parasitology/*methods ; RNA, Helminth/genetics ; RNA, Ribosomal, 5S/genetics ; Real-Time Polymerase Chain Reaction/*methods ; Sensitivity and Specificity ; Transition Temperature ; Wuchereria bancrofti/classification/genetics/*isolation & purification

C2H2 zinc-finger motif presents in 3% of proteins that are encoded in the human genome, and has the abilities to recognize DNA, RNA and protein. With nearly 3 decades of efforts, the mechanisms of zinc-finger mediated biomolecule recognitions have been studied to various extents. Zinc-finger binds into the major groove of DNA double helix, establishes an one-to-one recognition format between DNA bases and certain amino acids in a zinc-finger, and achieves specificity based on DNA sequences. While RNA molecules show a large variety in their structures, zinc-finger recognizes RNA through the collected information of specially displayed bases and special backbone folding. Initial studies have been performed on zinc-finger mediated protein-protein interactions. Existing data indicate multiple recognition modes. The studies on molecular mechanism have supported the development of engineered zinc-fingers, which have been introduced into applications. For its wide existence, large functional diversity and potential in translational applications, zinc-finger deserves a systematic study in every aspect.
Amino Acid Sequence ; Animals ; Binding Sites ; DNA ; chemistry ; genetics ; Humans ; Ikaros Transcription Factor ; chemistry ; genetics ; Nuclear Proteins ; chemistry ; genetics ; Protein Binding ; Proteins ; chemistry ; genetics ; RNA, Ribosomal, 5S ; chemistry ; genetics ; Transcription Factor TFIIIA ; chemistry ; genetics ; Transcription Factors ; chemistry ; genetics ; Vesicular Transport Proteins ; chemistry ; genetics ; Zinc Fingers