E111 | Pvu II, 2000
(1) A Mutant Sumo Facilitates Quick Plasmid Construction for Expressing Proteins with Native N-termini After Tag Removal.[TOP]Pubmed ID :28349302
Publication Date : 2017/03/28
Sumo is one of the fusion tags commonly used to enhance the expression and the solubility of recombinant proteins. One advantage of using sumo is that the removal of the sumo tag is highly specific because its recognition by a sumo protease is determined by its structural characteristics, instead of the sequence of a short peptide. Recently, it was reported that sumo could also be used as a protease recognition site to facilitate the removal of other fusion tags, such as MBP, when sumo itself is not suitable to enhance the solubility of a particular target protein. Using sumo as a recognition site is highly desirable when the target protein needs to have its native N terminus. However, constructing such a plasmid involves more than one cloning step because the N terminus of the target protein needs to be the next residue after the diglycine of sumo. Here, we report the construction of a new vector with a mutant sumo tag. The incorporation of a Pvu II site near the 3' end of tag coding sequence enables quick construction of plasmids for producing proteins with native termini. Its usage includes producing recombinant food allergens for studying conformational IgE epitopes.
Authors : Zhang Yuzhu, Fan Yuting,
(2) Comparative assessment of "plaque/media" change on three modalities of IVUS immediately after implantation of either everolimus-eluting bioresorbable vascular scaffold or everolimus-eluting metallic stent in Absorb II study.[TOP]Pubmed ID :28012050
Publication Date : 2016/12/24
The purpose of the study to assess the comparability of immediate changes in plaque/media volume (PV) on three modalities of intravascular ultrasound (IVUS) after implantation of either bioresorbable vascular scaffold (BVS) or everolimus-eluting metallic stent (EES) in Absorb II Study. The two devices have different device volume and ultrasound backscattering that may interfere with the "plaque/media" assessed by three modalities on IVUS: grayscale, backscattering of radiofrequency and brightness function. In a multicenter randomized controlled trial, 501 patients with stable or unstable angina underwent documentary IVUS pre- and post- implantation. The change in plaque/media volume (PV) was categorized into three groups according to the relative PV change in device segment: PV "increased" >+5% (PVI), PV unchanged ±5% (PVU), and PV decreased <-5% (PVD). The change in PV was re-evaluated three times: after subtraction of theoretical device volume, after analysis of echogenicity based on brightness function. In 449 patients, 483 lesions were analyzed pre- and post-implantation. "PVI" was more frequently observed in BVS (53.8%) than EES group (39.4%), p = 0.006. After subtraction of the theoretical device volume, the frequency of "PVI" decreased in both BVS (36.2%) and EES (32.1%) groups and became comparable (p = 0.581). In addition, the percentage of "PVI" was further reduced in both device groups after correction for either radiofrequency backscattering (BVS 34.4% vs. EES 22.6%) or echogenicity (BVS 25.2% vs. EES 9.7%). PV change in device segment was differently affected by BVS and EES devices implantation due to their differences in device volume and ultrasound backscattering. It implies that the lumen volume was also artifactually affected by the type of device implanted. Comparative IVUS assessment of lumen and plaque/media volume changes following implantation of BVS and EES requires specific methodological adjustment.
Authors : Zeng Yaping, Cavalcante Rafael, Tenekecioglu Erhan, Suwannasom Pannipa, Sotomi Yohei, Collet Carlos, Abdelghani Mahammad, Jonker Hans, Digne Franck, Horstkotte Dieter, Zehender Manfred, Indolfi Ciro, Saia Francesco, Fiorilli Rosario, Chevalier Bernard, Bolognese Leonardo, Goicolea Javier, Nie Shaoping, Onuma Yoshinobu, Serruys Patrick W, ,
(3) Estrogen receptor gene polymorphism in patients with degenerative lumbar scoliosis.[TOP]Pubmed ID :27399961
Publication Date : 2016/07/11
To examine the association between development of degenerative lumbar scoliosis (DLS) and sex hormones.
Authors : Park Yang Soo, Suh Kuen Tak, Shin Jong Ki, Lee Jung Sub,
(4) A Rapid and Sensitive Detection of Aflatoxin-producing Fungus Using an Optimized Polymerase Chain Reaction (PCR).[TOP]Pubmed ID :26977262
Publication Date : 2016/03/15
Aflatoxin B1 (AFB1) is produced by Aspergillus flavus growing in feedstuffs. Early detection of maize contamination by aflatoxigenic fungi is advantageous since aflatoxins exert adverse health effects. In this study, we report the development of an optimized conventional PCR for AFB1 detection and a rapid, sensitive and simple screening Real-time PCR (qPCR) with SYBR Green and two pairs of primers targeting the aflR genes which involved aflatoxin biosynthesis. AFB1 contaminated maize samples were divided into three groups by the toxin concentration. Genomic DNA was extracted from those samples. The target genes for A. flavus were tested by conventional PCR and the PCR products were analyzed by electrophoresis. A conventional PCR was carried out as nested PCR to verify the gene amplicon sizes. PCR-RFLP patterns, obtained with Hinc II and Pvu II enzyme analysis showed the differences to distinguish aflatoxin-producing fungi. However, they are not quantitative and need a separation of the products on gel and their visualization under UV light. On the other hand, qPCR facilitates the monitoring of the reaction as it progresses. It does not require post-PCR handling, which reduces the risk of cross-contamination and handling errors. It results in a much faster throughout. We found that the optimal primer annealing temperature was 65°C. The optimized template and primer concentration were 1.5 μL (50 ng/μL) and 3 μL (10 μM/μL) respectively. SYBR Green qPCR of four genes demonstrated amplification curves and melting peaks for tub1, afIM, afIR, and afID genes are at 88.0°C, 87.5°C, 83.5°C, and 89.5°C respectively. Consequently, it was found that the four primers had elevated annealing temperatures, nevertheless it is desirable since it enhances the DNA binding specificity of the dye. New qPCR protocol could be employed for the determination of aflatoxin content in feedstuff samples.
Authors : Bintvihok Anong, Treebonmuang Supitchaya, Srisakwattana Kitiya, Nuanchun Wisut, Patthanachai Koranis, Usawang Sungworn,
(5) Genotyping of Mycobacterium avium subsp. avium isolates from naturally infected lofts of domestic pigeons in Ahvaz by IS901 RFLP.[TOP]Pubmed ID :26719782
Publication Date : 2015/12/31
Avian tuberculosis is one of the most important infections affecting most species of birds. Mycobacterium avium can not only infect all species of birds, but also infect some domesticated mammals. The most crucial aspect of control and eradication scheme is identification of infection sources and transmission routs. Molecular techniques such as restriction fragment length polymorphism and pulse field gel electrophoresis have been shown to be much more discriminatory and suitable for use in the epidemiological study.
Authors : Parvandar-Asadollahi Kaveh, Mosavari Nader, Mayahi Mansoor,
(6) Polymorphisms in the gene encoding estrogen receptor alpha are associated with osteoarthritis in Han Chinese women.[TOP]Pubmed ID :25664105
Publication Date : 2015/02/09
Polymorphisms in the Xba I and Pvu II restriction enzyme recognition sites in the estrogen receptor-alpha gene (ESR1) have been associated with multiple diseases, including osteoarthritis. To determine whether such polymorphisms are associated with osteoarthritis in a Han Chinese population, 98 women with osteoarthritis and 196 healthy women were genotyped by PCR-RFLP of ESR1 with Xba I and Pvu II. Absence of a restriction polymorphism is indicated as an X or P allele; presence of the restriction polymorphism is indicated as an x or p allele. Clinical information was collected on each participant, including body weight, body mass index (BMI), knee radiograms, and bone mineral density (BMD). Body weight and BMI were higher for each Xba I genotype (all P < 0.05) in individuals with osteoarthritis compared to controls (p < 0.05). Femoral BMD was also significantly higher in the osteoarthritis group (p < 0.05). Additionally, the xx genotype for ESR1 was a significant risk factor for osteoarthritis (OR=1.98, 95% CI: 1.13~4.20, p=0.036). Thus, consistent with findings in other populations, the estrogen receptor genotype xx appears to be associated with susceptibility to osteoarthritis among Han Chinese women.
Authors : Liu Wei, Shao Feng-Min, Yan Lei, Cao Hui-Xia, Qiu Dong,
(7) Role of 3 lipoprotein lipase variants in triglycerides in children receiving highly active antiretroviral therapy.[TOP]Pubmed ID :24988117
Publication Date : 2015/03/09
Lipoprotein lipase is a key enzyme in lipid metabolism, especially for plasma triglycerides (TGs). Genetic variants have been associated with lipid levels in healthy individuals, cardiovascular disease, obesity and diabetes. Our aim was to evaluate the influence of 3 polymorphisms: Hind III, Pvu II and S447X in plasma TG levels in human immunodeficiency virus-1-infected children under highly active antiretroviral therapy (HAART).
Authors : Colombero Cecilia, Catano Gabriel, Rocco Carlos A, Mecikovsky Débora, Bologna Rosa, Aulicino Paula C, Sen Luisa, Mangano Andrea,
(8) Interaction of genetic risk factors confers increased risk for metabolic syndrome: the role of peroxisome proliferator-activated receptor γ.[TOP]Pubmed ID :24200052
Publication Date : 2014/01/09
The aim of the study was to estimate the influence of interactions between peroxisome proliferator-activated receptor γ (PPARγ) and target genes lipoprotein lipase (LPL), interleukin 6 (IL6), angiotensin converting enzyme (ACE), and angiotensin II type 1 receptor (AT1R) on metabolic syndrome (MetSy) and its traits.
Authors : Božina Tamara, Sertić Jadranka, Lovrić Jasna, Jelaković Bojan, Šimić Iveta, Reiner Željko,
(9) IS6110 restriction fragment length polymorphism typing of drug-resistant Mycobacterium tuberculosis strains from northeast South Africa.[TOP]Pubmed ID :23617199
Publication Date : 2013/04/26
Tuberculosis (TB) remains a deadly infectious disease affecting millions of people worldwide; 95% of TB cases, with 98% of death occur in developing countries. The situation in South Africa merits special attention. A total of 21,913 sputum specimens of suspected TB patients from three provinces of South Africa routinely submitted to the TB laboratory of Dr. George Mukhari (DGM) Hospital were assayed for Mycobacterium tuberculosis (MTB) growth and antibiotic susceptibility. The genetic diversity of 338 resistant strains were also studied. DNA isolated from the strains were restricted with Pvu II, transferred on to a nylon membrane and hybridized with a PCR-amplified horseradish peroxidase 245 bp IS6110 probe. Of the 338 resistant strains, 2.09% had less than 5 bands of IS6110, and 98% had 5 or more bands. Unique restriction fragment length polymorphism (RFLP) patterns were observed in 84.3% of the strains, showing their epidemiological independence, and 15.7% were grouped into 22 clusters. Thirty-two strains (61.5%) from the 52 that clustered were from Mpumalanga, 16/52 (30.8%) from Gauteng, and 4/52 (9.6%) from Limpopo province. Clustering was not associated with age. However, strains from male patients in Mpumalanga were more likely to be clustered than strains from male patients in Limpopo and/or Gauteng province. The minimum estimate for the proportion of resistant TB that was due to transmission is 9.06% (52-22 = 30/331). Our results indicate that transmission of drug-resistant strains may contribute substantially to the emergence of drug-resistant tuberculosis in South Africa.
Authors : Green Ezekiel, Obi Lawrence C, Okoh Anthony I, Nchabeleng Maphoshane, de Villiers Babsie E, Letsoalo Tomas, Hoosen Anwar A, Bessong Pascal O, Ndip Roland N,
(10) [Allele genotype analysis of Plasmodium vivax merozoite surface protein 1 (PvMSP-1) from Dangdong, Liaoning Province].[TOP]Pubmed ID :23072145
Publication Date : 2012/10/17
Nested PCR method was used to amplify the Plasmodium vivax merozoite surface protein 1 (PvMSP-1) gene fragment containing the ICB5 and ICB6 region from Plasmodium vivax in Liaoning Province. The PCR products were digested by Pvu II restriction endonuclease and the digested fragments were observed by 1.5% agarose gel electrophoresis, and all followed by sequencing analysis and comparison. In 11 field isolates of P. vivax, two kinds of DNA fragments with 470 and 400 bp were produced respectively. After PvuII digestion, two Sal-1 type fragments (120 and 350 bp) were obtained from 5 samples of 470 bp. Single band of 400 bp appeared in 1 samples as Belem type. Two bands of 120 and 280 bp appeared from another 1 sample as recombination type III, and other 4 bands with 120 and 240 bp as Korean isolate. The principal types of PvMSP-1 alleles exist in malaria endemic areas in Liaoning Province with no mixed infection of two different type alleles.