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Molecular cloning

Xiaofen Zhang, Guoyun Wang, Bin Chen, Heshan Du, Fenglan Zhang, Haiying Zhang, Qian Wang, Sansheng Geng
First flower node (FFN) is an important trait for evaluating fruit earliness in pepper (Capsicum annuum L.), but the genetic mechanisms that control FFN are still poorly understood. In the present study, we developed 249 F2 plants derived from an intraspecific cross between the inbred pepper lines Z4 and Z5. Thirty plants with the highest FFN and 30 plants with the lowest FFN were chosen and their DNAs were pooled according to phenotype to construct two bulked DNA pools. Specific-locus amplified fragment sequencing (SLAF-seq) was combined with bulked segregant analysis (BSA) to identify candidate regions related to FFN...
2018: PloS One
Jingting Shu, Gaige Ji, Ming Zhang, Yunjie Tu, Yanju Shan, Yifan Liu, Xiaojun Ju, Di Zhang
TNNI1 encodes the slow skeletal muscle isoform of troponin I. In the present study, the basic characteristic and expressing profile of the TNNI1 gene was first explored in Gaoyou ducks. Full-length TNNI1 cDNA of Gaoyou duck was obtained using reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE). The cDNA consisted of a 57-base pair (bp) 5'UTR, a 345-bp 3'UTR, and a 564-bp open reading frame. The predicted protein was predicted to be hydrophilic, nonsecretory protein and contained 17 phosphorylation sites...
March 20, 2018: Animal Biotechnology
Qingxia Yao, Karl P Fischer, D Lorne Tyrrell, Klaus S Gutfreund
Programmed death-1 (PD-1), upon engagement by its ligands, programmed death ligand-1 (PD-L1) and programmed death ligand-2 (PD-L2), provides signals that attenuate adaptive immune responses. Here we describe the identification of the Pekin duck PD-L2 (duPD-L2) and its gene structure. The duPD-L2 cDNA encodes a 321 amino acid protein that has an amino acid identity of 76% and 35% with chicken and human PD-L2, respectively. Mapping of the duPD-L2 cDNA with duck genomic sequences revealed an exonic structure similar to that of the human P dcd1lg2 gene...
March 2018: Biochemistry and Biophysics Reports
Qi Wang, Qian Wang, Yan-Le Zhou, Shuang Shan, Huan-Huan Cui, Yong Xiao, Kun Dong, Adel Khashaveh, Liang Sun, Yong-Jun Zhang
The phytophagous mirid bugs of Apolygus lucorum, Lygus pratensis as well as three Adelphocoris spp., including Adelphocoris lineolatus, A. suturalis , and A. fasciaticollis are major pests of multiple agricultural crops in China, which have distinct geographical distribution and occurrence ranges. Like many insect species, these bugs heavily rely on olfactory cues to search preferred host plants, thereby investigation on functional co-evolution and divergence of olfactory genes seems to be necessary and is of great interest...
2018: Frontiers in Physiology
Tom Verbiest, Rosemary Finnon, Natalie Brown, Lourdes Cruz-Garcia, Paul Finnon, Grainne O'Brien, Eleanor Ross, Simon Bouffler, Cheryl L Scudamore, Christophe Badie
Epidemiological studies have demonstrated an increased leukemia incidence following ionizing radiation exposure, but to date, the target cells and underlying mechanisms of radiation leukemogenesis remain largely unidentified. We engineered a mouse model carrying a different fluorescent marker on each chromosome 2, located inside the minimum deleted region occurring after radiation exposure and recognized as the first leukemogenic event. Using this tailored model, we report that following radiation exposure, more than half of asymptomatic CBA Sfpi1GFP/mCh mice presented with expanding clones of preleukemic hematopoietic cells harboring a hemizygous interstitial deletion of chromosome 2...
March 3, 2018: Leukemia: Official Journal of the Leukemia Society of America, Leukemia Research Fund, U.K
Bahare Salehi, Hossein Goudarzi, Bahram Nikmanesh, Hamidreza Houri, Mostafa Alavi-Moghaddam, Zohreh Ghalavand
RATIONALE: Acinetobacter baumannii is one of the antibiotic-resistant superbugs that threatens hospitalized patients. Emergence and spread of the multidrug-resistant (MDR) and extensively drug-resistant (XDR) clones cause erratic outbreaks following environmental contamination of hospital settings. OBJECTIVE: The present study intended to characterize the antimicrobial resistant profiles and the genotypes of clinical and environmental isolates of A. baumannii as a result of dissemination of resistant strains...
March 16, 2018: Journal of Infection and Chemotherapy: Official Journal of the Japan Society of Chemotherapy
Renata N Florindo, Valquiria P Souza, Lívia R Manzine, Cesar M Camilo, Sandro R Marana, Igor Polikarpov, Alessandro S Nascimento
Bifidobacterium is an important genus of probiotic bacteria colonizing the human gut. These bacteria can uptake oligosaccharides for the fermentative metabolism of hexoses and pentoses, producing lactate, acetate as well as short-chain fatty acids and propionate. These end-products are known to have important effects on human health. β-glucosidases (EC are pivotal enzymes for the metabolism and homeostasis of Bifidobacterium, since they hydrolyze small and soluble saccharides, typically producing glucose...
March 16, 2018: Biochimie
Norma Oviedo, Leticia Manuel-Apolinar, Sandra Orozco-Suárez, Teresa Juárez-Cedillo, Vilma Carolina Bekker Méndez, Emiliano Tesoro-Cruz
BACKGROUND: Intranasal administration (Int adm) has been well-studied and offers the possibility to deliver larger molecular weight biologics, such as proteins, viral vectors, nanoparticles, and naked plasmids to the brain and treat a variety of diseases in the central nervous system. The predominant challenge in this field is finding efficient vectors that are capable of crossing the blood-brain barrier (BBB). OBJECTIVES: Here, we investigated whether a naked plasmid (pIRES-hrGFP-1a), could cross the BBB, reach brain cells and express green fluorescent protein (GFP) after int-adm and propose it as candidate for future gene therapy studies...
March 16, 2018: Archives of Medical Research
Zhideng Lin, Yisheng Huang, Weiguang Zou, Hua Rong, Meiling Hao, Xiaobo Wen
Enzymes that lengthen the carbon chain of polyunsaturated fatty acids (PUFA) are key to the biosynthesis of the long-chain polyunsaturated fatty acids (LC-PUFA). Here we report on the molecular cloning, tissue distribution, functional characterization and nutritional regulation of a elovl5 gene from Nibea coibor. The full-length cDNA was 1315 bp, including a 5-untranslated region (UTR) of 134 bp, a 3-UTR of 296 bp and an open reading frame of 885 bp, which specified a peptide of 294 amino acids. Bioinformatics analysis showed that the deduced peptide sequence possessed all the characteristic features of microsomal fatty acyl elongases, including the so-called histidine box (HXXHH), the canonical C-terminal endoplasmic reticulum retention signal, several predicted transmembrane regions and other highly conserved motifs...
March 16, 2018: Gene
Philip J Pedersen, Kirsten B Thomsen, Jon B Flak, Maria A Tejada, Frank Hauser, Dagmar Trachsel, Rikke Buhl, Theodore Kalbfleisch, Michael Scott DePriest, James N MacLeod, Kirstine Calloe, Dan A Klaerke
No abstract text is available yet for this article.
March 16, 2018: Research in Veterinary Science
LiLi Shi, Bin Li, Ting Ting Zhou, Wei Wang, Siuming F Chan
The recent use of RNA-Seq to study the transcriptomes of different species has helped identify a large number of new genes from different non-model organisms. In this study, five distinctive transcripts encoding for neuropeptide members of the CHH/MIH/GIH family have been identified from the spermatophore transcriptome of the shrimp Fenneropenaeus merguiensis. The size of these transcripts ranged from 531 bp to 1771 bp. Four transcripts encoded different CHH-family subtype I members, and one transcript encoded a subtype II member...
2018: PloS One
Monica M Lahra, Irene Martin, Walter Demczuk, Amy V Jennison, Ken-Ichi Lee, Shu-Ichi Nakayama, Brigitte Lefebvre, Jean Longtin, Alison Ward, Michael R Mulvey, Teodora Wi, Makoto Ohnishi, David Whiley
Ceftriaxone remains a first-line treatment for patients infected by Neisseria gonorrhoeae in most settings. We investigated the possible spread of a ceftriaxone-resistant FC428 N. gonorrhoeae clone in Japan after recent isolation of similar strains in Denmark (GK124) and Canada (47707). We report 2 instances of the FC428 clone in Australia in heterosexual men traveling from Asia. Our bioinformatic analyses included core single-nucleotide variation phylogeny and in silico molecular typing; phylogenetic analysis showed close genetic relatedness among all 5 isolates...
April 2018: Emerging Infectious Diseases
Sunil Ghatge, Youri Yang, Woo-Young Song, Tae-Young Kim, Hor-Gil Hur
In the present study, the gene encoding a multicopper oxidase, more precisely a laccase from the thermoalkaliphilic aerobic bacterium Caldalkalibacillus thermarum strain TA2.A1 (CtLac), was cloned and expressed in Escherichia coli. CtLac is a monomeric protein with a molecular weight of 57 kDa as determined by native polyacrylamide gel electrophoresis. The optimum pH and temperature for 2,6-dimethoxyphenol (2,6-DMP) oxidation were 8.0 and 70 °C, respectively. The kinetic constants Km and kcat for 2,6-DMP were of 200 μM and 23 s-1 , respectively...
March 19, 2018: Applied Microbiology and Biotechnology
Mohammed Sami Alhaddad, Afnan Khalifah AlBarjas, Lolowah Ebraheem Alhammar, Abdullatif Sami Al Rashed, Lorina Ineta Badger-Emeka
Background: Acinetobacter baumannii , is an emerging nosocomial multidrug resistance pathogen with the rapid spread of clones being reported in health-care settings and hospitals worldwide. Carbapenem resistance in this bacterium has been attributed to D OXA β-lactamases with OXA-51-like β-lactamase, being present in all A. baumannii isolate. The present study looks into the antibiotics susceptibility and molecular characterization of clinical A. baumannii isolates from Intensive Care Unit (ICU) samples in Al-Hofuf, South-eastern region of Saudi Arabia...
January 2018: International Journal of Applied and Basic Medical Research
Jun Bai, Bin-Jian Yang, Xin Luo
5-hydroxy-4'-nitro-7-propionyloxy-genistein (HNPG), a novel synthetic isoflavone derivative, was demonstrated to possess antitumor activity in gastric cancer and breast cancer in vitro , but its antitumor effect and mechanism in ovarian cancer has not been characterized. The aim of the present study was to investigate the effects of HNPG on inhibiting the proliferation and invasion in human ovarian cancer A2780 cell lines of cisplatin resistance (A2780/DDP) and elucidate its underlying molecular mechanism. The results indicated that HNPG presented with marked antitumor activity against A2780/DDP cells in vitro , significantly inhibited the rates of proliferation, clone formation, invasion and metastasis, and markedly induced apoptosis in dose- and time-dependent manner...
April 2018: Oncology Letters
Tong Xie, Lingzhao Mo, Li Li, Naiquan Mao, Danrong Li, Deseng Liu, Chuantian Zuo, Dingming Huang, Qi Pan, Li Yang, Shoufeng Wang
The present study aimed to isolate and characterize side population (SP) cells in the human lung cancer A549 cell line, and elucidate the molecular mechanism of SP cells underlying lung cancer. The SP and non-SP (NSP) cells in A549 cells were isolated and their differentiation was analyzed by fluorescence-activated cell sorting. An in vitro plate clone assay, Matrigel® Transwell assay and chemoresistance analysis of the sorted SP and NSP cells were performed. In addition, the sorted SP and NSP cells were injected into BALB/c nude mice to detect their tumorigenic potential in vivo ...
April 2018: Oncology Letters
Liang-Qing Dong, Yang Shi, Li-Jie Ma, Liu-Xiao Yang, Xiao-Ying Wang, Shu Zhang, Zhi-Chao Wang, Meng Duan, Zhao Zhang, Long-Zi Liu, Bo-Hao Zheng, Zhen-Bin Ding, Ai-Wu Ke, Da-Ming Gao, Ke Yuan, Jian Zhou, Jia Fan, Ruibin Xi, Qiang Gao
BACKGROUND & AIMS: Intrahepatic cholangiocarcinoma (ICC) is the second-most lethal primary liver cancer. Little is known about intratumoral heterogeneity (ITH) and its impact on ICC progression. We aim to investigate its ITH in hope of helping develop new therapeutic strategies. METHODS: We obtained 69 spatially distinct regions from 6 operable ICCs. Patient-derived primary cancer cells (PDPCs) were established for each region, followed by whole-exome sequencing(WES) and multi-level validation...
March 15, 2018: Journal of Hepatology
Jesse J McClure, Xiaoyang Li, C James Chou
Since the identification and cloning of human histone deacetylases (HDACs) and the rapid approval of vorinostat (Zolinza®) for the treatment of cutaneous T-cell lymphoma, the field of HDAC biology has met many initial successes. However, many challenges remain due to the complexity involved in the lysine posttranslational modifications, epigenetic transcription regulation, and nonepigenetic cellular signaling cascades. In this chapter, we will: review the discovery of the first HDAC inhibitor and present discussion regarding the future of next-generation HDAC inhibitors, give an overview of different classes of HDACs and their differences in lysine deacylation activity, discuss different classes of HDAC inhibitors and their HDAC isozyme preferences, and review HDAC inhibitors' preclinical studies, their clinical trials, their pharmacokinetic challenges, and future direction...
2018: Advances in Cancer Research
Pengfei Chu, Libo He, Lv Xiong, Lifei Luo, Rong Huang, Lanjie Liao, Yongming Li, Zuoyan Zhu, Yaping Wang
The mammalian sterile 20-like (MST) family, which belongs to the serine/threonine protein kinase superfamily, has five members that can be found in mammals: STK3 (also called MST2), STK4 (MST1), STK24 (MST3), STK25 (YSK1 or SOK1), and STK26 (MST4). The MST kinases have key roles in apoptosis, immune regulation, inflammatory responses, cancer, and cell proliferation in mammals, whereas the roles and transcriptional regulatory mechanism of these kinases in teleost fish are still unclear. In this study, four STK genes (CiSTK3, CiSTK24, CiSTK25, and CiSTK26) were cloned and analyzed in grass carp (Ctenopharyngodon idella)...
March 14, 2018: Fish & Shellfish Immunology
Xiqi Li, Cesar A Arias, Samuel L Aitken, Jessica Galloway Peña, Diana Panesso, Michael Chang, Lorena Diaz, Rafael Rios, Yazan Numan, Sammi Ghaoui, Sruti DebRoy, Micah M Bhatti, Dawn E Simmons, Isaam Raad, Ray Hachem, Stephanie A Folan, Pranoti Sahasarabhojane, Awdhesh Kalia, Samuel A Shelburne
Background: Pathobionts, bacteria that are typically human commensals but can cause disease, contribute significantly to antimicrobial resistance. Staphylococcus epidermidis is a prototypical pathobiont as it is a ubiquitous human commensal but also a leading cause of healthcare-associated bacteremia. We sought to determine the etiology of a recent increase in invasive S. epidermidis isolates resistant to linezolid. Methods: Whole-genome sequencing (WGS) was performed on 176 S...
March 12, 2018: Clinical Infectious Diseases: An Official Publication of the Infectious Diseases Society of America
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