Identification of novel transcripts and sRNA of Brucella melitensis by RNA-Seq / 中国人兽共患病学报

To identify novel transcripts and sRNA in genome of B .melitensis by transcriptome sequencing ,total RNA were extracted from B .melitensis culture and rRNA were removed .After the addition of adaptor ,RNA was reversely transcribed into cDNA ,which were then subjected to PCR amplification and sequencing .The generated reads were mapped to genome se‐quence of B .melitensis strain 16M .With the mapping results ,novel transcripts and sRNA were identified by bioinformatics methods .Sequencing results analysis showed that genome sequence was covered with the reads with good quality .A total of 773 genes were extended in their 5′and/or 3′ends of their original locations .Sixteen novel transcripts and 241 sRNAs candi‐dates were identified .RT‐PCR showed that some of the sRNAs were differentially expressed under stress conditions .In B . melitensis genome ,there is novel transcript which is not predicted .The sRNA does exist in B .melitensis and were expressed under different conditions .

Transcriptional analysis of Brucella virulence regulation genes under stress conditions and during cell infection / 中华微生物学和免疫学杂志

Objective To study the important virulence regulation genes of Brucella,and to understand their function.Methods Quantitative RT-PCR was used to quantify their relative transcription profiles under stress conditions and during macrophage cell infection.Results These genes were activated at different levels under these conditions and during cell infection,indicating their roles in pathogenesis at different srage of infection.Conclusion The transcription profiles of these genes have different effects about their functions.

Detection and Genotyping of Herps Simplex Virus Type 2 by DNA Sequencing / 解放军医学杂志

Objective Herpes simplex virus is the pathogenic agent of human herpes simplex. There are two genotypes of herpes simplex virus, HSV-1 and HSV-2. The clinical manifestations of HSV-1 and HSV-2 overlap each other and it is difficult to differentiate them clinically. Methods developed based on genome differences are efficient ones to differentiate the two genotypes of HSV. In this study, we attempted to develop a new method to detection and genotyping human HSV in clinical samples. Methods Swab samples were collected from genital lesions of patients and placed in transport media. Samples were used to inoculate Vero cells. After appearance of the cytotoxicity, the infection mixtures were collected, and subjected to genomic DNA extraction. Based on the conservation and variation of gD of HSV-1 and HSV-2, a pair of primers amplifying both of them were designed and synthesized. Sequence of the virus were amplified and cloned into pMD-18T, and then the sequence was determined by DNA sequencing. The sequence was compared to all the known sequences in Genebank by using BLAST. According to the BLAST results, the genus and genotype of the virus was determined. Results A DNA fragment of about 200 bp was successfully amplified. This DNA fragment was cloned and sequenced. The sequence was compared with other known sequences. the results showed that this sequence had the highest homology to gD of HSV, indicating that the virus in the sample was HSV-2. The BLAST results also showed that there were some differences in the sequence of gD to those previously isolated. Conclusion DNA sequencing of PCR amplification products is an efficient and definite method to detect and genotype HSV-1 and HSV-2 which otherwise are difficult to differentiate clinically.