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@#Abstract: With the development of molecular biology, non-coding sRNA has been found to play an important regulatory role in gene expression and protein activity, affecting various biological pathways including mosquito resistance against insecticides. Understanding the molecular regulatory mechanisms of drug resistance is essential for controlling mosquitoes, , of which metabolic resistance being the most critical mechanism, mainly referring to the high expression of metabolic detoxification enzyme-related genes (especially the cytochrome P450 enzyme system) in mosquitoes. On the basis of verification of insecticide resistance-related genes, further research on the correlation between sRNA and mosquito resistance-related genes provides new ideas and directions for further exploring the mechanism of mosquito resistance. The study of mosquito metabolic resistance mechanism is of great significance for the control of vector mosquitoes, drug resistance monitoring and novel insecticide development. This article reviews the progress of research on the resistance genes, sRNAs biosynthesis, genes involved in regulating mosquito metabolic detoxification enzymes and their applications.
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ObjectiveQuorum-sensing (QS) and small regulatory RNA (sRNA) play key regulatory roles in many signaling cascades of Pseudomonas aeruginosa. To investigate whether sRNA is involved in P. aeruginosa QS system, screening QS system-related sRNA, and to construct sRNA overexpression and deletion strains of Pseudomonas aeruginosa for further study of sRNA function.MethodsSRNA associated with the QS system was screened by qPCR and RNA-sequencing (RNA-seq). The target gene were amplified by PCR and inserted into the overexpression vector pROp200 or the homologous recombination vector pGSM-MR, respectively. The connection reaction solution of pROp200-sRNA and pGSM-ΔsRNA was transformed into Escherichia coli DH5a and SM10lp, respectively. The recombinant vectors were identified by PCR. The pROp200-sRNA was transformed into PAO1 by heat shock method, and the pGSM-ΔsRNA was transferred from SM10lp to PAO1 by conjugation. SRNA overexpression and deletion strains were identified by PCR, DNA sequencing and qPCR, the determination of the growth curves and the pyocyanin levels of strains.ResultsFive QS -associated sRNA P26, P5316.1, P30, P34 and AmiL were successfully screened by RNA-seq and qPCR. PCR, DNA sequencing and qPCR showed that sRNA of AmiL, P30 and P34 overexpression and knockout were successful. Compared with wild-type strain, sRNA overexpression and knockout had no significant effect on bacterial growth curve. It were notably that overexpression of AmiL and P30 inhibited and increase the production of pyocyanin, respectively (P0.05).ConclusionThe sRNA overexpression and deletion strains have been successfully constructed and can be used to study the regulatory relationship between sRNA and QS systems, and to further functional study.
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Exosomes are nanoscale vesicles secreted by plant cells, containing DNA, small RNA(sRNA), microRNA(miRNA), protein and other substances and mediating cell-cell communication. miRNA is a highly efficient gene expression regulator, which can regulate cross-border gene expression. sRNA transfers between host and organism through plasma-desmata and induces gene silencing. Plant exosomes exert anti-inflammatory, anti-virus, anti-fibrosis, anti-tumor and other related effects, via the substances contained, and participate in the defense response of pathogen invasion. Most plant exosomes are edible and can be used as carriers to deliver specific drugs, with no toxicity and side effects. Plant exosomes have become a hotspot of researches. This paper summarizes and analyzes the progress in the plant exosomes research reported in the literature in recent years, focusing on the source and function of medicinal plant exosomes, so as to provide reference for future studies on plant exosomes.
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To obtain the microbial composition of traditional Chinese medicine of Faeces Trogopterori, ten samples were collected from the imitate wildness farmland in Shangluo City, Shaanxi Province. In this study, 16S rRNA gene was used as molecular marker to explore the microbiome and the sequences were analyzed by Usearch analysis platform. The COG and KEGG database is used to predict and analyze the function of the flora. A great number of 285 218 high quality clean reads with a length of 400-450 bp were obtained from 10 samples. Bacterial species detected in these samples covered 8 phyla, 25 families, 75 genera and 120 species. The dominant phylum microbial communities in these samples were Firmicutes (87.68% ± 2.68%) and the Bacteroidetes (7.62% ± 3.74%), all samples showed a high microbial diversity, the predicted functional metagenome was heavily involved in energy metabolism. This study provided that the beneficial bacteria in Faeces Trogopterori may be one of its active ingredients, and no pathogens are detected in the sample.
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Bacterial biofilm refers to a tunicate-like biological group composed of polysaccharide, protein and nucleic acid secreted by bacteria on the surface of the mucous membrane or biological material. The biofilm formation is a major cause of chronic infections. Bacteria could produce some secondary metabolites during the growth and reproduction. Some of them act as signaling molecules allowing bacteria to communicate and regulate many important physiological behaviors at multiple-cell level, such as bioluminescence, biofilm formation, motility and lifestyles. Usually, these signal molecules play an important role in the formation of bacterial biofilm. We review here the effects of related signal molecules of Quorum Sensing, cyclic diguanylate, Two-Component Systems and sRNA on the biofilm formation. Focusing on these regulation mechanism of signal molecules in the process of biofilm formation is necessary for the prevention and treatment of some chronic diseases.
Subject(s)
Bacterial Proteins , Biofilms , Cyclic GMP , Gene Expression Regulation, Bacterial , Protein Binding , Quorum SensingABSTRACT
Small RNAs(sRNAs) play a significant role in the regulation of bacterial growth.When sensing certain environmental cues such as fluctuation of nutrient concentration, temperature, pH, and osmolarity, sRNAs can influence the expression of target genes.The formation of biofilms is initiated by bacteria transitioning from the planktonic to the surface-associated mode of growth, which is a self-produced extracellular matrix composed of proteins, polysaccharides, and DNA.Recent evidences have shown that small RNA plays an important role in the regulation of bacterial biofilm formation.sRNAs have key roles in biofilm formation process by base pairing with target mRNAs or interaction with modulating proteins.This review discussed the regulation mechanism and pathway of sRNAs in bacterial biofilms formation, and summarized three classical regulatory models of sRNAs in bacterial biofilms formation, this review also gives the research status and development direction of sRNAs in bacterial biofilms formation.
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Objective: To detect the differential expression profile of miRNAs (microRNAs) in crude drugs and processed products of Lycopodiastrum casuarinoides and identify potential bioactive herbal-derived miRNAs. Methods: The samples of the whole aerial tissues (including stems, leaves, branches) were collected in Yichun area of Jiangxi Province and were authenticated by relevant experts. General RNA was extracted from the crude drugs and processed products of L. casuarinoides, respectively. High-qualified small RNAs (sRNA) were isolated to be constructed the sRNA sequencing library. Then, the single-ended sequencing was conducted by using Illumina HiSeqTM 2500 sequencing method. MiRNA characteristic of L. casuarinoides was analyzed by relevant bioinformatics. Results: 9 898 332 and 10 099 918 clean reads were obtained from crude drugs group and processed products group, respectively. A total of 25 microRNAs were differentially expressed between crude drugs group and processed products groups with statistical significance, among which 22 were up regulated and three were down-regulated in processed products group compared to those in the crude drugs group. GO (gene ontology) analysis showed that their homo sapiens targets were enriched in catalytic binding, molecular transducer and KEGG (Kyoto encyclopedia of genes and genomes) analysis was shown for the target genes enriched in cancer and immunity-related pathways, such as pathways in cancer, proteoglycans in cancer. Conclusion: The differential expression profile of microRNAs is first revealed by two processing forms of L. casuarinoides. This study suggests that the crude drugs and processed products miRNAs identified in this study will lay a foundation for exploring the pharmaceutical function of miRNAs in L. casuarinoides.
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Objective To study the profiles and function of small RNA (sRNA)gene during chondrogenesis in rats so as to clarify the mechanisms of chondrocytes proliferation and differentiation.Methods All the sRNAs were identified from the female SD rats femoral head cartilages at three time points:at birth,ablactation and maturation,and three sRNA libraries were constructed.The Solexa sequencing and the bioinformatics analysis were employed to be blasted with the genomes of SD rats.Results The perfect match reads in the three libraries were screened out,which were correspondent to the 21 7 921 (41.23%),1 96 650 (38.74%)and 245 436 (41.54%)unique sRNA sequence,respectively.The percentages of 20-24 nt sRNA were 71.94% (d0),72.85% (d21),and 86.39%(d42).Half of clean sequences were 22 nt sRNA.The distribution characteristics of the reads were in line with the high-quality sRNA.More than 62% clean reads were from mature miRNA while the ratios in the three libraries were only 0.69%,0.78% and 0.63%.About 60% of the unique sRNA could not be matched with miRBase20.0 or Rfam9.1.Conclusion The distribution model of miRNA in the three libraries indicates that the miRNAs with different functions or from different sources are involved in the regulation of chondrocytes proliferation and differentiation in bone development and formation.
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Objective To identify small non-coding RNAs encoded by plasmid pPCP1 and investigate their roles in biofilm formation, stress tolerance and/or virulence in Yersinia pestis.Methods Seven plasmid pPCP1-encoded sRNAs were identified by RNA-seq results in Y.pestis in our previous studies.Northern blot was used to validate the presence of the seven sRNAs.The sRNA-deletion mutants were constructed via λ-Red homologous recombination system.The biofilm formation, high salt tolerance and virulence of the phenotypes were compared between Y.pestis WT strain and sRNA mutants.Results and Conclusion The expression of seven pPCP1-encoded sRNAs was validated and the transcript length detected by Northern blotting corresponded to the length observed by RNA-seq.On this basis, five sRNA-deletion mutants were obtained.The capacity of biofilm formation was weakened upon deletion of sR3446.The tolerance of sR3446, sR3457, sR4338 and sR4340 mutants was found weakened in vitro compared to that of wild-type strain,but the tolerance of sR6143 was found increased.Slight virulence attenuation was found in two sRNA mutants ( sR4338 and sR4340 ) .The results suggest that pPCP1-deriving sRNA might be implicated in stress response, biofilm and virulence in Y.pestis.
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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 .
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Objective To conduct a pilot study on genome-wide in vivo protein-RNA interactions in E.coli.Methods Bacterial lysate was treated with RNase before the RNA fragments protected by proteins were extracted from treated lysate and used to construct cDNA library that was applied to high-throughput sequencing .Finally, the transcripts bound by proteins were obtained by bioinformatics analysis .Results A total of 3193 transcripts were obtained , including 2234 mRNAs, 47 sRNAs, 39 tRNAs, 11 rRNAs, and 862 intergenic regions .Conclusion Some information of transcripts interacting with proteins in E.coli is acquired , which will facilitate further studies of protein-RNA interactions .
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Small non-coding RNA (sRNA) is a kind of newly discovered 50 nt~500 nt small RNAs that do not encode proteins. To date, more than 150 sRNA have been found in bacteria. The small RNA acting by base-pairing with target mRNAs, resulting in post-transcriptionally regulating gene expression, is important regulators in the bacterial response to stress, virulence and metabolism. At present, researches of sRNA mainly based on bioinformatical prediction and molecular biological experiments. The sRNA that obtained through these methods needs confirmation in laboratory, and then study of its functions through a variety of experimental methods.