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Molecular cloning

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https://www.readbyqxmd.com/read/28230773/programmed-death-ligand-1-pd-l1-tumor-expression-is-associated-with-a-better-prognosis-and-diabetic-disease-in-triple-negative-breast-cancer-patients
#1
Gerardo Botti, Francesca Collina, Giosuè Scognamiglio, Federica Rao, Valentina Peluso, Rossella De Cecio, Michela Piezzo, Gabriella Landi, Michelino De Laurentiis, Monica Cantile, Maurizio Di Bonito
Triple Negative Breast Cancers (TNBC) subtype is an aggressive disease with poor clinical outcome. The only treatment available is surgery followed by chemotherapy or radiotherapy. Programmed death-ligand 1 (PD-L1) is a trans-membrane protein expressed on a wide variety of cells including immune cells, epithelial and vascular endothelial cells. Recently, PD-1/PD-L1 pathway signaling was described as an adaptive immune resistance mechanism enacted by the tumor cells to evade the immune response. Its presence on tumor cell membranes, acquired for this reason, through time, is an important prognostic value...
February 21, 2017: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/28229970/molecular-analysis-of-the-human-faecal-archaea-in-a-southern-indian-population
#2
Sandya B Rani, Ramadass Balamurugan, Balakrishnan S Ramakrishna
Archaea are an important constituent of the human gut microbiota, but there is no information on human gut archaea in an Indian population. In this study, faecal samples were obtained from different age groups (neonatal babies, preschool children, school-going children, adolescents, adults and elderly) of a southern Indian population, and from a tribal population also resident in southern India). 16S rRNA gene sequences specific to Archaea were amplified from pooled faecal DNA in each group, sequenced, and aligned against the NCBI database...
March 2017: Journal of Biosciences
https://www.readbyqxmd.com/read/28224997/clonal-reversal-of-ageing-associated-stem-cell-lineage-bias-via-a-pluripotent-intermediate
#3
Martin Wahlestedt, Eva Erlandsson, Trine Kristiansen, Rong Lu, Cord Brakebusch, Irving L Weissman, Joan Yuan, Javier Martin-Gonzalez, David Bryder
Ageing associates with significant alterations in somatic/adult stem cells and therapies to counteract these might have profound benefits for health. In the blood, haematopoietic stem cell (HSC) ageing is linked to several functional shortcomings. However, besides the recent realization that individual HSCs might be preset differentially already from young age, HSCs might also age asynchronously. Evaluating the prospects for HSC rejuvenation therefore ultimately requires approaching those HSCs that are functionally affected by age...
February 22, 2017: Nature Communications
https://www.readbyqxmd.com/read/28223988/the-impact-of-genotyping-by-sequencing-pipelines-on-snp-discovery-and-identification-of-markers-associated-with-verticillium-wilt-resistance-in-autotetraploid-alfalfa-medicago-sativa-l
#4
Long-Xi Yu, Ping Zheng, Suresh Bhamidimarri, Xiang-Ping Liu, Dorie Main
Verticillium wilt (VW) of alfalfa is a soilborne disease causing severe yield loss in alfalfa. To identify molecular markers associated with VW resistance, we used an integrated framework of genome-wide association study (GWAS) with high-throughput genotyping by sequencing (GBS) to identify loci associated with VW resistance in an F1 full-sib alfalfa population. Phenotyping was performed using manual inoculation of the pathogen to cloned plants of each individual and disease severity was scored using a standard scale...
2017: Frontiers in Plant Science
https://www.readbyqxmd.com/read/28223458/mutation-of-the-n-terminal-region-of-chikungunya-virus-capsid-protein-implications-for-vaccine-design
#5
Adam Taylor, Xiang Liu, Ali Zaid, Lucas Y H Goh, Jody Hobson-Peters, Roy A Hall, Andres Merits, Suresh Mahalingam
Mosquito-transmitted chikungunya virus (CHIKV) is an arthritogenic alphavirus of the Togaviridae family responsible for frequent outbreaks of arthritic disease in humans. Capsid protein, a structural protein encoded by the CHIKV RNA genome, is able to translocate to the host cell nucleolus. In encephalitic alphaviruses, nuclear translocation induces host cell transcriptional shutoff; however, the role of capsid protein nucleolar localization in arthritogenic alphaviruses remains unclear. Using recombinant enhanced green fluorescent protein (EGFP)-tagged expression constructs and CHIKV infectious clones, we describe a nucleolar localization sequence (NoLS) in the N-terminal region of capsid protein, previously uncharacterized in CHIKV...
February 21, 2017: MBio
https://www.readbyqxmd.com/read/28223161/molecular-characterization-of-pacific-oyster-crassostrea-gigas-irak4-gene-and-its-role-in-myd88-dependent-pathway
#6
Xueying Tang, Baoyu Huang, Linlin Zhang, Li Li, Guofan Zhang
Interleukin-1 receptor-associated kinases (IRAKs) play important roles in MyD88-dependent TLR signaling, the crucial innate immune pathway in molluscs. In this study, we examined the full-length IRAK4 genetic sequence in the Pacific oyster (Crassostrea gigas) by molecular cloning. Phylogenetic analysis revealed that CgIRAK4 is most closely related to Mytilus edulis, and forms a clade with other molluscs. CgIRAK4 transcripts are widely expressed in all tissues, with the highest expression observed in the hemocytes and gill...
February 13, 2017: Developmental and Comparative Immunology
https://www.readbyqxmd.com/read/28223110/h2a-and-ca-l-hipposin-gene-characteristic-analysis-and-expression-responses-to-aeromonas-hydrophila-infection-in-carassius-aurutus
#7
Xianghui Kong, Xiangmin Wu, Chao Pei, Jie Zhang, Xianliang Zhao, Li Li, Guoxing Nie, Xuejun Li
Antimicrobial peptide is an important component of the host innate immune system and thus serves a crucial function in host defense against microbial invasion. In this study, H2A and derived antimicrobial peptide Ca-L-hipposin were cloned and characterized in Carassius aurutus. The gene H2A full-length cDNA is 908 bp and includes a 5'-terminal untranslated region (UTR) of 55 bp and a 3'-terminal UTR of 466 bp with a canonical polyadenylation signal sequence AATAA, as well as an open reading frame (ORF) of 387 bp encoding a polypeptide of 128 amino acids, with a molecular weight of 13...
February 18, 2017: Fish & Shellfish Immunology
https://www.readbyqxmd.com/read/28223030/heterologous-expression-of-a-rice-metallothionein-isoform-osmti-1b-in-saccharomyces-cerevisiae-enhances-cadmium-hydrogen-peroxide-and-ethanol-tolerance
#8
Zahra Ansarypour, Azar Shahpiri
Metallothioneins are a superfamily of low-molecular-weight, cysteine (Cys)-rich proteins that are believed to play important roles in protection against metal toxicity and oxidative stress. The main purpose of this study was to investigate the effect of heterologous expression of a rice metallothionein isoform (OsMTI-1b) on the tolerance of Saccharomyces cerevisiae to Cd(2+), H2O2 and ethanol stress. The gene encoding OsMTI-1b was cloned into p426GPD as a yeast expression vector. The new construct was transformed to competent cells of S...
February 4, 2017: Brazilian Journal of Microbiology: [publication of the Brazilian Society for Microbiology]
https://www.readbyqxmd.com/read/28222377/identification-of-autophagy-related-genes-lc3-and-atg4-from-yellow-catfish-pelteobagrus-fulvidraco-and-their-transcriptional-responses-to-waterborne-and-dietborne-zinc-exposure
#9
Chuan-Chuan Wei, Zhi Luo, Yu-Feng Song, Ya-Xiong Pan, Kun Wu, Wen-Jing You
Autophagy mediates the regulation of lipid metabolism. Moreover, our recent study indicated that waterborne and dietborne zinc (Zn) exposure differentially influenced lipid metabolism in a fish species of significance for aquaculture, yellow catfish Pelteobagrus fulvidraco, but further mechanism remained unknown. The hypothesis of the present study is that autophagy mediated the Zn-induced changes of lipid metabolism of yellow catfish subjected to different exposure pathways. To this end, we cloned key genes involved in autophagy in yellow catfish, explored their mRNA expressions in responses to different Zn exposure pathways...
February 8, 2017: Chemosphere
https://www.readbyqxmd.com/read/28221880/spread-of-a-major-clone-of-salmonella-enterica-serotype-enteritidis-in-poultry-and-in-salmonellosis-outbreaks-in-southern-brazil
#10
Karen Apellanis Borges, Thales Quedi Furian, Sara Neves de Souza, Eduardo César Tondo, André Felipe Streck, Carlos Tadeu Pippi Salle, Hamilton Luiz de Souza Moraes, Vladimir Pinheiro do Nascimento
Salmonella spp. are among the most important agents of foodborne diseases all over the world. Human Salmonella outbreaks are often associated with the consumption of poultry products (meat and eggs), and one of the most prevalent serotypes associated with these products is Salmonella Enteritidis. Brazil is one of the most important poultry exporters in the world. In southern Brazil, three closely related clones of Salmonella Enteritidis have been responsible for the majority of foodborne Salmonella outbreaks over the past decade...
January 2017: Journal of Food Protection
https://www.readbyqxmd.com/read/28220547/identification-and-molecular-epidemiology-of-dermatophyte-isolates-by-repetitive-sequence-pcr-based-dna-fingerprinting-using-the-diversilab-system-in-turkey
#11
A Nedret Koc, Mustafa A Atalay, Melek Inci, Fatma M Sariguzel, Hafize Sav
Dermatophyte species, isolation and identification in clinical samples are still difficult and take a long time. The identification and molecular epidemiology of dermatophytes commonly isolated in a clinical laboratory in Turkey by repetitive sequence-based PCR (rep-PCR) were assessed by comparing the results with those of reference identification. A total of 44 dermatophytes isolated from various clinical specimens of 20 patients with superficial mycoses in Kayseri and 24 patients in Hatay were studied. The identification of dermatophyte isolates was based on the reference identification and rep-PCR using the DiversiLab System (BioMerieux)...
February 21, 2017: Mycoses
https://www.readbyqxmd.com/read/28220435/rapid-construction-of-multiplexed-crispr-cas9-systems-for-plant-genome-editing
#12
Levi Lowder, Aimee Malzahn, Yiping Qi
Multiplex CRISPR-Cas9 nuclease mediated genome editing is an efficient method for disrupting gene function in plants. Use of CRISPR-Cas9 has escalated rapidly in recent years and is expected to become routine practice in molecular biology and related fields of research. Due to the relatively novel and widespread adoption of this technology, first-time users may not have regular access to experienced guidance or technical support from peers or mentors. Here, we offer guidance and technical support in the form of a detailed and tested protocol for simultaneous targeting of three separate loci on the TRANSPARENT TESTA 4 (TT4) gene in Arabidopsis thaliana using multiplex CRISPR-Cas9...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28220352/purification-characterization-and-overexpression-of-an-endo-1-4-%C3%AE-mannanase-from-thermotolerant-bacillus-sp-swu60
#13
Weeranuch Seesom, Polphet Thongket, Tomohiro Yamamoto, Shigeo Takenaka, Tatsuji Sakamoto, Wasana Sukhumsirichart
Endo-β-1,4-mannanases are important catalytic agents in several industries. The enzymes randomly cleave the β-1,4-linkage in the mannan backbone and release short β-1,4-mannooligosaccharides and mannose. In the present study, mannanase (ManS2) from thermotolerant Bacillus sp. SWU60 was purified, characterized, and its gene was cloned and overexpressed in Escherichia coli. ManS2 was purified from culture filtrate (300 ml) by using hydrophobic, ion-exchange, and size-exclusive liquid chromatography. The apparent molecular mass was 38 kDa...
March 2017: World Journal of Microbiology & Biotechnology
https://www.readbyqxmd.com/read/28219197/-down-regulation-of-mir-146b-5p-promotes-malignant-transformation-of-fusion-cells-after-co-culture-of-macrophages-with-glioma-stem-cells-in-vitro
#14
H H Cai, H Y Wang, H R Liu, Y J Sheng, D G Xi, Y P Xue, X L Dai, A D Wang, Q Huang, J Dong
Objective: To observe mutual interactions between macrophages(Mφ) and glioma stem cells (GSCs)in dual-color tracing model in vitro, to identify the biological characteristics of fusion cells in multiple levels, and to analysis the relevant molecular mechanisms. Methods: Red fluorescent protein(RFP) gene was stably transfected into human GSCs cell line SU4. Mφ cells were obtained from Balb/c nude mice with enhanced green fluorescent protein (EGFP) expression. Then two cells were co-cultured in dual-color tracing platform...
February 7, 2017: Zhonghua Yi Xue za Zhi [Chinese medical journal]
https://www.readbyqxmd.com/read/28219109/the-use-of-braf-v600e-mutation-specific-immunohistochemistry-in-pediatric-langerhans-cell-histiocytosis
#15
Leomar Y Ballester, Miguel D Cantu, Karen P H Lim, Stephen F Sarabia, Lizmery Suarez Ferguson, C Renee Webb, Carl E Allen, Kenneth L McClain, Carrie A Mohila, Jyotinder N Punia, Angshumoy Roy, Dolores H López-Terrada, M John Hicks, Kevin E Fisher
BRAF p.V600E mutations are detected in greater than 50% of pediatric Langerhans cell histiocytosis (LCH) lesions. However, the use of mutation-specific BRAF V600E immunohistochemistry (IHC) as a surrogate for molecular testing in pediatric LCH is unknown. We tested the mutation-specific BRAF V600E monoclonal antibody (clone VE1) in formalin-fixed, paraffin-embedded LCH samples from 26 pediatric patients (14 males and 12 females, ages 7 mo-17 y) using allele-specific real-time polymerase chain reaction (PCR) with a limit of detection of 0...
February 20, 2017: Hematological Oncology
https://www.readbyqxmd.com/read/28218974/molecular-cloning-bioinformatics-analysis-and-transcriptional-profiling-of-jaz1-and-jaz2-from-salvia-miltiorrhiza
#16
Yangyun Zhou, Xun Zhou, Qing Li, Junfeng Chen, Ying Xiao, Lei Zhang, Wansheng Chen
Production of major effective metabolites, tanshinones and lithospermic acid B (LAB), was dramatically enhanced by exogenous jasmonate (JA) treatment in Salvia miltiorrhiza. However, the molecular mechanism of such metabolic activation in S. miltiorrhiza has not been elucidated yet. Here, we focused on jasmonate ZIM-domain (JAZ) proteins that act as repressors of JA signaling. Open reading frames of two novel genes, SmJAZ1 and SmJAZ2, from S. miltiorrhiza were amplified according to the annotation of S. miltiorrhiza transcriptome...
January 2017: Biotechnology and Applied Biochemistry
https://www.readbyqxmd.com/read/28218427/two-vitellogenins-in-the-loliginid-squid-uroteuthis-edulis-identification-and-specific-expression-in-ovarian-follicles
#17
Hajime Kitano, Naoki Nagano, Keishi Sakaguchi, Michiya Matsuyama
Vitellogenenesis is a physiological process common in oviparous animals. The molecular profile, modifications, and utilization of vitellogenin (VTG), a precursor of yolk protein, have been characterized in various taxa to understand oogenesis within different modes of reproduction. Hormonal regulation of VTGs has been investigated in invertebrates, such as insects and crustaceans; conversely, little is known for cephalopods. In this study, we isolated two VTG genes (ue-VTG1 and ue-VTG2) from the loliginid swordtip squid, Uroteuthis edulis, via a comprehensive survey of a transcriptome database and subsequent cDNA cloning...
February 20, 2017: Molecular Reproduction and Development
https://www.readbyqxmd.com/read/28217858/golden-gate-assembly-system-dedicated-to-complex-pathway-manipulation-in-yarrowia-lipolytica
#18
Ewelina Celińska, Rodrigo Ledesma-Amaro, Macarena Larroude, Tristan Rossignol, Cyrille Pauthenier, Jean-Marc Nicaud
In this study, we have adopted Golden Gate modular cloning strategy to develop a robust and versatile DNA assembly platform for the nonconventional yeast Yarrowia lipolytica. To this end, a broad set of destination vectors and interchangeable building blocks have been constructed. The DNA modules were assembled on a scaffold of predesigned 4 nt overhangs covering three transcription units (each bearing promoter, gene and terminator), selection marker gene and genomic integration targeting sequences, constituting altogether thirteen elements...
February 19, 2017: Microbial Biotechnology
https://www.readbyqxmd.com/read/28216998/genetic-mapping-of-yield-traits-using-ril-population-derived-from-fuchuan-dahuasheng-and-icg6375-of-peanut-arachis-hypogaea-l
#19
Yuning Chen, Xiaoping Ren, Yanli Zheng, Xiaojing Zhou, Li Huang, Liying Yan, Yongqing Jiao, Weigang Chen, Shunmou Huang, Liyun Wan, Yong Lei, Boshou Liao, Dongxin Huai, Wenhui Wei, Huifang Jiang
The genetic architecture determinants of yield traits in peanut (Arachis hypogaea L.) are poorly understood. In the present study, an effort was made to map quantitative trait loci (QTLs) for yield traits using recombinant inbred lines (RIL). A genetic linkage map was constructed containing 609 loci, covering a total of 1557.48 cM with an average distance of 2.56 cM between adjacent markers. The present map exhibited good collinearity with the physical map of diploid species of Arachis. Ninety-two repeatable QTLs were identified for 11 traits including height of main stem, total branching number, and nine pod- and seed-related traits...
2017: Molecular Breeding: New Strategies in Plant Improvement
https://www.readbyqxmd.com/read/28216323/cdna-cloning-and-expression-analysis-of-a-phosphopyruvate-hydratase-gene-from-the-chinese-shrimp-fenneropenaeus-chinensis
#20
Xupeng Li, Xianhong Meng, Kun Luo, Sheng Luan, Baoxiang Cao, Jie Kong
In the present study a cDNA encoding a phosphopyruvate hydratase (enolase) was cloned from the muscle of the Chinese shrimp (Fenneropenaeus chinensis) and named as FcEnolase. The cDNA of FcEnolase encoded a protein of 434 amino acid residues with a molecular mass 47.22 kDa. The residues 342-355 constituted the signature motif "LLLKVNQIGSVTES". A SNP locus (C96T) in the ORF at 96 bp was identified. The results showed that the FcEnolase was a conserved gene. In the normal F. chinensis, the mRNA level in the muscle was much higher (P < 0...
February 13, 2017: Fish & Shellfish Immunology
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