1.Idiopathic Spinal Epidural Lipomatosis.
Journal of Korean Neurosurgical Society 2001;30(6):795-799
Spinal epidural lipomatosis, which causes symptomatic compression of neural elements, is a relatively uncommon disease. Although it has been reported frequently in association with the administration of exogenous steroids, a few cases of epidural lipomatosis with no association to exogenous steroids, have been reported. Idiopathic spinal epidural lipomatosis may be a separate disease from that induced by steroid. Here, the authors present two cases of symptomatic epidural lipomatosis with no history of steroid-dependent diseases and review the relevant literature.
DNA Transposable Elements
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Lipomatosis*
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Steroids
2.A novel resolution vector with Bacillus thuringiensis plasmid replicon ori44.
Lan WU ; Ming SUN ; Chen-Guang ZHU ; Lei ZHANG ; Zi-Niu YU
Chinese Journal of Biotechnology 2002;18(3):335-338
The resolution recognization sites of transposon Tn4430 of Bacillus thuringiensis was inserted into cloning vector pRSET B and pUC19, resulting recombinant plasmids pBMB1201 and pBMB1202. Both of the mini res fragments, BamHI/HindIII fragment in pBMB1201 and EcoRI/HindIII fragment in pBMB1202, were ligated to the 3.3 kb EcoRI/HindIII fragment of shuttle vector pHT3101, which contained the ori. Ec, ampr and emr antibiotic resistant genes, resulting recombinant plasmid pBMB1203. After deleted the BamHI and EcoRI sites which located ouside the two res sites, resolution vector pBMB1204 was resulted. There are multiple cloning sites between two copies of resolution sites which have the same direction. The plasmid replication origin ori44, which come from B. thuringiensis sub sp. kurstaki strain YBT-1520, was inserted into the multiple cloning sites of pBMB1204 and then resolution shuttle vector pBMB1205 was obtained. With spectinomycin resistant gene as target, it was found that the resolution rate is 100% and the stability of the resolved plasmid is 93%. Using this shuttle vector, antibiotic resistance markers and other non-B. thuringiensis DNA can be selectively eliminated after the selection of transformants by antibiotic resistance marker. This vector is very useful to solve the gene safety problem while has no effect on target gene expression.
Bacillus thuringiensis
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genetics
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DNA Transposable Elements
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Genetic Vectors
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Plasmids
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Replicon
3.Dissemination of insertion sequences IS605, IS606 among clinical isolates of Helicobacter pylori in China.
Maojun ZHANG ; Jianzhong ZHANG ; Lihua HE ; Haoyan GUO ; Yan YIN ; Zengfen ZHOU
Chinese Journal of Epidemiology 2002;23(5):366-369
OBJECTIVETo study the distribution of IS605, IS606 among clinical isolates of Helicobacter pylori in China.
METHODSA total of 104 H.pylori strains isolated from 5 different geographic regions in China were analyzed by PCR and dot-blot.
RESULTSForty-two strains out of the 104 isolates from 5 regions in China were found containing IS605 with 19 containing IS606. The frequency (66%) of IS605 positive strains from Yunnan province was higher than that from other areas. The different distribution of IS606 was neither associated with geographical regions nor with the presence of IS605 but IS606 were associated with the different clinical outcomes. However, the two reading frames ORFA and ORFB of IS605 were constantly coexisting.
CONCLUSIONIn China, IS605 and IS606 of H. pylori were widely existing but the presence of IS605 in H. pylori might be associated with geographic origin.
DNA Transposable Elements ; Helicobacter pylori ; genetics ; Humans ; Polymerase Chain Reaction
4.Comparison of MITE transposons mPing in different rice subspecies.
Ning ZHANG ; Yanan RUAN ; Shanshan WANG ; Yang LIU ; Chen ZHAO ; Jingjing WANG ; Kaixi WANG ; Yanli WANG ; Hongyan WANG
Chinese Journal of Biotechnology 2016;32(9):1264-1272
The mPing family is the first active MITE TE family identified in rice genome. In order to compare the compositions and distributions of mPing family in the genomes of two rice subspecies japonica (cv. Nipponbare) and indica (cv. 93-11), we initially estimated the copy numbers of mPing family in those two subspecies using Southern blot and then confirmed the results by searching homologous copies in each reference genome using Blastn program, which turned out to have 52 and 14 mPing copies in corresponding reference genome, respectively. All mPing members in Nipponbare genome belong to mPing-1, while there are 3 mPing-1 and 11 mPing-2 copies in 93-11 genome. By further investigating the 5-kb flanking sequences of those mPing copies, it was found that 23 and 3 protein-coding genes in Nipponbare and 93-11 genome are residing adjacent to those mPing copies respectively. These results establish the preliminary theoretical foundation for further dissecting the genetic differences of japonica and indica rice in terms of the diversities and distributions of their component mPing.
Animals
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DNA Transposable Elements
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genetics
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Genome, Plant
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Oryza
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classification
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genetics
5.Recent advances in CRISPR-related transposable elements.
Shuqing NING ; Xinxin WU ; Yunzi LUO
Chinese Journal of Biotechnology 2022;38(12):4371-4384
A new wave of research has been inspired by the CRISPR-Cas system with respect to their application in genome editing. The CRISPR-Cas system can not only be applied in gene knockout and insertion, but also be used in base editing, transcriptional regulation and recombination of gene clusters. However, the low efficiency of homology-directed repair (HDR) limits its application. Unlike the CRISPR-Cas system, mobile genetic elements (MGE) can insert DNA fragments into cell chromosomes without the aid of HDR. Recently, it is reported that CRISPR-related transposable elements can guide targeted DNA insertion. Their transposition mechanisms and reprogramming abilities have brought novel opportunities to the development of this field. This review summarized the research progress and application development of natural CRISPR-related transposable elements in recent years, as well as the applications of fused dCas9-transposase. It proposed the application prospects and potential challenges of CRISPR-related transposable elements in the future, which provided a reference for the development direction of gene editing tools.
DNA Transposable Elements/genetics*
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Gene Editing
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CRISPR-Cas Systems/genetics*
6.Relationship between drug resistance of Pseudomonas aeruginosa isolated from burn wounds and its mobile genetic elements.
Xi-Hao HU ; Xiao-Min XU ; Zu-Huang MI ; You-Fen FAN ; Wei-Yun FENG
Chinese Journal of Burns 2009;25(2):103-105
OBJECTIVETo investigate the relationship between the drug resistance of Pseudomonas aeruginosa (PA) isolated from burn patients wounds and its mobile genetic elements, including plasmid, transposon, and integron.
METHODSThirty-two strains of PA were isolated from wounds exudate of hospitalized burn patients in Ningbo No. 2 Hospital. PA drug sensitivity was determined using GNS-448 drug sensitivity card and K-B tests. The genetic markers of plasmid, transposon and integron including traA, traF, tnpA, tnpU, merA, int I 1 were amplified by PCR and verified by gene sequencing.
RESULTSDrug resistant rate of 32 PA strains to gentamicin, amikacin, cefoperazone/sulbactam, ciprofloxacin was 43.7%, 32.0%, 46.8%, 49.9%, respectively. PA drug resistant rates to piperacillin, cefotaxime, ceftazidime, cefepime, aztreonam, piperacillin/tazobactam, levofloxacin, imipenem and meropenem were all above 56.0%. Seventeen out of 32 PA strains were found to carry transposon and (or) integron genetic markers. One strain was positive for both tnpA and merA, 8 strains were positive for both merA and int I 1, 1 strain was only positive for tnpA, 2 strains were only positive for merA, and 5 strains were positive for int I 1 only.
CONCLUSIONSPA isolated from burn wounds of hospitalized patients in Ningbo No. 2 Hospital is seriously drug resistant, which may relate with its high positive rate of mobile genetic elements of transposon and (or) integron.
Anti-Bacterial Agents ; pharmacology ; Burns ; microbiology ; DNA Transposable Elements ; Drug Resistance, Multiple, Bacterial ; genetics ; Humans ; Integrons ; Microbial Sensitivity Tests ; Plasmids ; Pseudomonas aeruginosa ; drug effects ; genetics ; isolation & purification
7.Foldback Intercoil DNA and the Mechanism of DNA Transposition.
Genomics & Informatics 2014;12(3):80-86
Foldback intercoil (FBI) DNA is formed by the folding back at one point of a non-helical parallel track of double-stranded DNA at as sharp as 180degrees and the intertwining of two double helixes within each other's major groove to form an intercoil with a diameter of 2.2 nm. FBI DNA has been suggested to mediate intra-molecular homologous recombination of a deletion and inversion. Inter-molecular homologous recombination, known as site-specific insertion, on the other hand, is mediated by the direct perpendicular approach of the FBI DNA tip, as the attP site, onto the target DNA, as the attB site. Transposition of DNA transposons involves the pairing of terminal inverted repeats and 5-7-bp tandem target duplication. FBI DNA configuration effectively explains simple as well as replicative transposition, along with the involvement of an enhancer element. The majority of diverse retrotransposable elements that employ a target site duplication mechanism is also suggested to follow the FBI DNA-mediated perpendicular insertion of the paired intercoil ends by non-homologous end-joining, together with gap filling. A genome-wide perspective of transposable elements in light of FBI DNA is discussed.
DNA End-Joining Repair
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DNA Transposable Elements
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DNA*
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Enhancer Elements, Genetic
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Hand
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Homologous Recombination
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Retroelements
8.Virulence genes of Agrobacterium tumefaciens and analysis function of its biology.
Xiao-Ping LU ; Ya-Xiang XU ; Kojima MINEO
Chinese Journal of Biotechnology 2003;19(6):651-654
Chromosomal virulence genes acvB, abvA, chvA of Agrobacterium tumefaciens were cloned with the technique of transposon 5 insertion. The chromosome genes are necessary for Agrobacterium tumfaciens absorbing to cell ular surface of plant, the adherence reaction can't be executed and result in losing the toxicity if mutations are occurred in some chromosome genes. The chromosome toxicity gene is inactivated due to transposon Tn5 be inserted and the accept ant cell infected with Agrobacterium tumefaciens can't cause tumor ultimately. This article briefly introduces the research way of thinking and strategy of this technique and the important roles of every gene, which are taken of in the process of T-DNA's form, transfer, integration, and expression etc. This article also gives a presumption to T-DNA's transport: The plant cell wall's porin may be T-DNA's natural channel.
Agrobacterium tumefaciens
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genetics
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metabolism
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pathogenicity
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DNA Transposable Elements
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genetics
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physiology
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Virulence
;
genetics
9.Modification and decoration of transposase: a review.
Chinese Journal of Biotechnology 2014;30(10):1504-1514
Transposons are the mobile and autonomic replication DNA fragments in genomes. With more understanding of the structure and function of transposons, numerous transposons have been developed to the genetics tool for gene function analysis, gene transformation and gene therapy. The low transpositional activity of the natural transposons is the main obstacles to the utilization of transposons. Recently, with the progress in bioinformatics and protein engineering methods, researchers have reconstructed and optimized natural transposases to create hyperactive transposases that catalyze the transposition with high efficiency. The resulted hyperactive transposons have been applied to gene-modification and gene-tagging. Meanwhile, transposase chimeras were created by protein fusion technology. The insertion characteristic of transposons were artificially regulated which could be utilized in gene therapy.
DNA Transposable Elements
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Gene Targeting
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Genetic Therapy
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Protein Engineering
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Transposases
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chemistry
10.Construction of Tn5 transposon insertion mutants of Ralstonia solanacearum isolated from Pogostemon cablin.
Ya-Qin WANG ; Yu-Yao ZHANG ; Hong HE ; Zhuan LI ; Zhi-Cheng DENG ; Hua JIN ; Guang-Wei LI
China Journal of Chinese Materia Medica 2019;44(1):77-81
Ralstonia solanacearum strain PRS-84 used in this study was isolated from diseased Pogostemon cablin plants in our previous study.The competent cells of R.solanacearum strain PRS-84 were transformed by electroporation with Tn5 transposon and then were plated on TTC agar plates containing kanamycin to select for kanamycin-resistant colonies.The detection of kanamycin-resistant gene in kanamycin-resistant colonies was performed by PCR.Further,the flanking fragments of Tn5 transposon insertion site in the mutants were amplified by inverse PCR,and the flanking fragments were sequenced and analyzed.The results indicated that the kanamycin-resistant colonies were obtained in the transformation experiment of R.solanacearum strain PRS-84 by electroporation with Tn5 transposon.A specific band of approximately 700 bp was amplified by PCR from kanamycin-resistant colonies.The flanking sequences of Tn5 transposon insertion site in the transformants were obtained by inverse PCR.After sequencing and sequence analysis of Tn5 transposon insertion site in mutants,we preliminarily speculated that the Tn5 transposon inserted in the typ A gene,rec O gene and gid A gene in three mutants,respectively.A random mutagenesis system of R.solanacearum strain PRS-84 by electroporation with Tn5 transposon has been established,and the Tn5 insertion mutants have been obtained.This study might facilitate the creation of mutant library and the discovery of the virulence gene of R.solanacearum isolated from P.cablin.
DNA Transposable Elements
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Electroporation
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Genes, Bacterial
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Mutagenesis, Insertional
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Pogostemon
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microbiology
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Ralstonia solanacearum
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genetics
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Virulence