scholarly journals Cloning of Acyl-ACP Thioesterase FatA fromArachis hypogaeaL. and Its Expression inEscherichia coli

2012 ◽  
Vol 2012 ◽  
pp. 1-5 ◽  
Author(s):  
Gao Chen ◽  
Zhen-ying Peng ◽  
Lei Shan ◽  
Ning Xuan ◽  
Gui-ying Tang ◽  
...  
Keyword(s):  
Oleic Acid ◽  
Palmitoleic Acid ◽  
Membrane Lipid ◽  
Pcr Analysis ◽  
Fatty Acid Profiles ◽  
Reading Frame ◽  
E Coli ◽  
Real Time Quantitative Pcr ◽  
Immature Seeds ◽  
Expression Inescherichia Coli

In this study, a full-length cDNA of the acyl-ACP thioesterase,AhFatA, was cloned from developing seeds ofArachis hypogaeaL. by 3′-RACE. Sequence analysis showed that the open reading frame encodes a peptide of 372 amino acids and has 50–70% identity with FatA from other plants. Real-time quantitative PCR analysis revealed thatAhFatA was expressed in all tissues ofA. hypogaeaL., but most strongly in the immature seeds harvested at 60 days after pegging. Heterologous expression ofAhFatA inEscherichia coliaffected bacterial growth and changed the fatty acid profiles of the membrane lipid, resulting in directed accumulation towards palmitoleic acid and oleic acid. These results indicate that AhFatA is at least partially responsible for determining the high palmitoleic acid and oleic acid composition ofE. coli.

scholarly journals Molecular cloning, expression and characterization of Bmserpin-2 gene from Bombyx mori.

2009 ◽  
Vol 56 (4) ◽  
Author(s):  
Ye Pan ◽  
Hengchuan Xia ◽  
Peng Lü ◽  
KePing Chen ◽  
Qin Yao ◽  
...  
Keyword(s):  
Bombyx Mori ◽  
Developmental Stages ◽  
Pcr Analysis ◽  
Post Inoculation ◽  
Amino Acid Residues ◽  
Gene Encoding ◽  
Reading Frame ◽  
E Coli ◽  
Real Time Quantitative Pcr ◽  
Localization Analysis

Serpins are a broadly distributed family of protease inhibitors. In this study, the gene encoding Bombyx mori serpin-2 (Bmserpin-2) was cloned and expressed in E. coli. The Bmserpin-2 cDNA contains a 1125 bp open reading frame (ORF). The deduced protein has 374 amino-acid residues, contains a conserved SERPIN domain and shares extensive homology with other invertebrate serpins. RT-PCR analysis showed that Bmserpin-2 was expressed in all developmental stages of B. mori larvae and various larval tissues. Subcellular localization analysis indicated that Bmserpin-2 protein was located in the cytoplasm. Interestingly, real-time quantitative PCR revealed that the expression of Bmserpin-2 in the midgut of susceptible B. mori strain 306 significantly increased at 72 hours post inoculation (hpi) when infected with BmNPV. However, there was no significant increase of the Bmserpin-2 expression in resistant strain NB infected with BmNPV. Thus, our data indicates that Bmserpin-2 may be involved in B. mori antiviral response.

scholarly journals Dissemination of CTX-M-Type β-Lactamases among Clinical Isolates of Enterobacteriaceae in Paris, France

2004 ◽  
Vol 48 (4) ◽  
pp. 1249-1255 ◽  
Author(s):  
C. Eckert ◽  
V. Gautier ◽  
M. Saladin-Allard ◽  
N. Hidri ◽  
C. Verdet ◽  
...  
Keyword(s):  
Insertion Sequence ◽  
Dna Fingerprint ◽  
Clinical Isolates ◽  
Pcr Analysis ◽  
Reading Frame ◽  
E Coli ◽  
Paris Area ◽  
The Family ◽  
Genes Encoding ◽  
Cephalosporin Resistance

ABSTRACT We analyzed 19 clinical isolates of the family Enterobacteriaceae (16 Escherichia coli isolates and 3 Klebsiella pneumoniae isolates) collected from four different hospitals in Paris, France, from 2000 to 2002. These strains had a particular extended-spectrum cephalosporin resistance profile characterized by a higher level of resistance to cefotaxime and aztreonam than to ceftazidime. The bla CTX-M genes encoding these β-lactamases were involved in this resistance, with a predominance of bla CTX-M-15. Ten of the 19 isolates produced both TEM-1- and CTX-M-type enzymes. One strain (E. coli TN13) expressed CMY-2, TEM-1, and CTX-M-14. bla CTX-M genes were found on large plasmids. In 15 cases the same insertion sequence, ISEcp1, was located upstream of the 5′ end of the bla CTX-M gene. In one case we identified an insertion sequence designated IS26. Examination of the other three bla CTX-M genes by cloning, sequencing, and PCR analysis revealed the presence of a complex sul1-type integron that includes open reading frame ORF513, which carries the bla gene and the surrounding DNA. Five isolates had the same plasmid DNA fingerprint, suggesting clonal dissemination of CTX-M-15-producing strains in the Paris area.

scholarly journals Monounsaturated Fatty Acids Are Substrates for Aldehyde Generation in Tellurite-ExposedEscherichia coli

2013 ◽  
Vol 2013 ◽  
pp. 1-7 ◽  
Author(s):  
Gonzalo A. Pradenas ◽  
Waldo A. Díaz-Vásquez ◽  
José M. Pérez-Donoso ◽  
Claudio C. Vásquez
Keyword(s):  
Oxidative Stress ◽  
Fatty Acid ◽  
Bacterial Resistance ◽  
Monounsaturated Fatty Acids ◽  
Membrane Lipid ◽  
Fatty Acid Profiles ◽  
Wild Type ◽  
E Coli ◽  
Membrane Oxidation ◽  
Cellular Components

Reactive oxygen species (ROS) damage macromolecules and cellular components in nearly all kinds of cells and often generate toxic intracellular byproducts. In this work, aldehyde generation derived from theEscherichia colimembrane oxidation as well as membrane fatty acid profiles, protein oxidation, and bacterial resistance to oxidative stress elicitors was evaluated. Studies included wild-type cells as well as cells exhibiting a modulated monounsaturated fatty acid (MUFA) ratio. The hydroxyaldehyde 4-hydroxy 2-nonenal was found to be most likely produced byE. coli, whose levels are dependent upon exposure to oxidative stress elicitors. Aldehyde amounts and markers of oxidative damage decreased upon exposure toE. colicontaining low MUFA ratios, which was paralleled by a concomitant increase in resistance to ROS-generating compounds. MUFAs ratio, lipid peroxidation, and aldehyde generation were found to be directly related; that is, the lower the MUFAs ratio, the lower the peroxide and aldehyde generation levels. These results provide additional evidence about MUFAs being targets for membrane lipid oxidation and their relevance in aldehyde generation.

scholarly journals Isolation, Characterization, and Expression Analysis of the MaMDH Gene in Banana

HortScience ◽  
2013 ◽  
Vol 48 (5) ◽  
pp. 614-619
Author(s):  
Cai-Hong Jia ◽  
Ju-Hua Liu ◽  
Zhi-Qiang Jin ◽  
Qiu-Ju Deng ◽  
Jian-Bin Zhang ◽  
...  
Keyword(s):  
Real Time ◽  
Quantitative Polymerase Chain Reaction ◽  
Expression Patterns ◽  
Ethylene Biosynthesis ◽  
Pcr Analysis ◽  
Malic Dehydrogenase ◽  
Reading Frame ◽  
Expression Levels ◽  
Post Harvest ◽  
Real Time Quantitative Pcr

A full-length cDNA isolated from banana (Musa acuminata L. AAA group) fruit was named MaMDH, containing an open reading frame encoding 332 amino acids that represents the gene for cytoplasmic malic dehydrogenase (MDH). Sequence analysis showed that MaMDH shares high similarity with MDHs from castor bean (XP_002533463), tobacco (CAC12826), peach (AAL11502), and chickpeas (CAC10208). Real-time quantitative polymerase chain reaction (PCR) analysis of MaMDH spatial expression showed that it was expressed in all organs examined: roots, rhizomes, leaves, flowers, and fruits. The expression was the highest in flowers followed by the fruits and roots, whereas the rhizomes and leaves displayed the lowest expression levels. Real-time quantitative PCR revealed that MaMDH exhibited differential expression patterns in post-harvest banana fruits correlating with ethylene biosynthesis. In naturally ripened banana fruits, MaMDH expression was in accordance with ethylene biosynthesis. In accordance, for banana fruits treated with the ethylene analog 1-methylclopropene (1-MCP), MaMDH expression levels were inhibited and remained constant. After treatment with ethylene, MaMDH expression in banana fruits significantly increased with ethylene biosynthesis and peaked 3 days after harvest, which was 11 days earlier than that in naturally ripened banana fruits. These results suggest that MaMDH expression is induced by ethylene to regulate post-harvest banana fruits ripening.

scholarly journals Cloning and Expression Analysis of Flavonoid 3′, 5′-Hydroxylase Gene from Brunfelsia acuminata

Genes ◽  
2021 ◽  
Vol 12 (7) ◽  
pp. 1086
Author(s):  
Min Li ◽  
Yuting Cao ◽  
Biswojit Debnath ◽  
Hongjuan Yang ◽  
Xiaohua Kui ◽  
...  
Keyword(s):  
Petunia Hybrida ◽  
Molecular Breeding ◽  
Flower Color ◽  
Cloning And Expression ◽  
Pcr Analysis ◽  
Blue Flower ◽  
Rt Pcr ◽  
Reading Frame ◽  
Real Time Quantitative Pcr ◽  
H Gene

The full-length cDNA sequence of F3′5′H gene from the Brunfelsia acuminata was obtained by RT-PCR and RACE, whose GenBank accession number is JQ678765. The sequence contains a 1521 bp open reading frame, 120 bp 5′UTR and 61 bp 3′UTR, encoding a total of 506 amino acids. The molecular mass of the predicted protein is 56.47 kDa with an estimated pI of 8.78, respectively. Sequence alignment showed that the amino acid sequence of F3′5′H was 91%, 87% and 84% with that of Petunia × hybrida, Nierembergia sp., Solanum tuberosum, respectively. Real-time quantitative PCR analysis showed that the expression of F3′5′H gene was different in petals of different days, which was the highest expression level on day 0 and significantly higher than other days. The results indicated that F3′5′H might play key role in flower color regulation and provide a theoretical reference for blue flower molecular breeding.

scholarly journals Determination of Seasonal Changes in the Fat and Fatty Acid Profiles of Saurida lessepsianus (Russell, Golani and Tikochinski, 2015) Caught from Mersin Bay

2019 ◽  
Vol 7 (9) ◽  
pp. 1483
Author(s):  
Mısra Bakan ◽  
Elif Ayşe Erdoğan Eliuz ◽  
Deniz Ayas
Keyword(s):  
Fatty Acids ◽  
Oleic Acid ◽  
Fatty Acid ◽  
Seasonal Changes ◽  
Palmitoleic Acid ◽  
Winter Season ◽  
Fatty Acid Profiles ◽  
Seasonal Factors ◽  
Autumn And Winter

In this study, seasonal changes in the lipid and fatty acid profiles of S. lessepsianus caught from the Mersin Bay were investigated. The total lipid levels of S. lessepsianus were found to be 2.94%, 7.19%, 2.45%, 0.83%, in spring, summer, autumn and winter season, respectively. Major fatty acids in S. lessepsianus were palmitic acid, stearic acid, oleic acid, palmitoleic acid, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) in all seasons. The highest values of palmitic, palmitoleic and EPA were determined as 22.97%, 3.80% and 4.22% in spring, respectively. The highest values of stearic and oleic acid were determined as 15.93% and 7.84% in autumn, respectively. The highest value of DHA were also determined as 31.91% in winter season. The EPA level from polyunsaturated fatty acids was found in the range of 2.54-4.22% (23.09-195.62 mg/100g). The highest level of DHA were observed in the winter season and its levels changed in the range of 19.83-31.81% and was calculated as 201.29-1301.73 mg/100g. In addition, the highest level of the Σn3, Σn6, and Σn9 were calculated in the summer season as 1516.39, 114.88, 399.77 mg/100g, respectively. This report showed that fat and fatty acid profiles of S. lessepsianus are quite influenced by seasonal factors.

scholarly journals PROFIL ASAM LEMAK MINYAK SAWIT SETELAH PROSES PENGGORENGAN IKAN

2018 ◽  
Vol 6 (1) ◽  
pp. 30
Author(s):  
Silvana Dinaintang Harikedua ◽  
Vera T Harikedua
Keyword(s):  
Oleic Acid ◽  
Fatty Acid ◽  
Palmitic Acid ◽  
Fatty Acid Profile ◽  
Palm Oil ◽  
Palmitoleic Acid ◽  
Major Fatty Acid ◽  
Fatty Acid Profiles ◽  
Frying Process

The fatty acid profile of palm oil is presented in this work. The palm oil (control) was rich in palmitic acid (39.01%) and oleic acid (44.50%). The results indicated that fish frying process for about 60 minutes was given little differences fatty acid profiles compared to control palm oil. The major fatty acid in the palm oil after frying fish was palmitic acid (41. 13%) and oleic acid (42.62), and developed the existenceof palmitoleic acid (0.28%), which is not found in control palm oil.

Molecular cloning and expression of a jasmonate biosynthetic gene allene oxide cyclase from Camellia sinensis

2016 ◽  
Vol 96 (1) ◽  
pp. 109-116 ◽  
Author(s):  
Meng-xin Wang ◽  
Qing-ping Ma ◽  
Bao-yu Han ◽  
Xing-hui Li
Keyword(s):  
Camellia Sinensis ◽  
Defense Responses ◽  
Tea Plant ◽  
Cloning And Expression ◽  
Pcr Analysis ◽  
Allene Oxide ◽  
Coding Region ◽  
Allene Oxide Cyclase ◽  
Reading Frame ◽  
Real Time Quantitative Pcr

As a family of signaling plant hormone, jasmonic acid plays an important role in coordinating plant defense responses to pests and pathogen attack through transcriptional and metabolic changes. In the jasmonate biosynthetic pathway of plants, allene oxide cyclase (AOC) is an essential enzyme. Here we cloned a cDNA from tea plant (Camellia sinensis), named as CsAOC (GenBank: HQ889679), which was 916 bp, containing an open reading frame (738 bp) encoding 245 amino acids. Comparative and bioinformatic analyses revealed that the deduced protein of CsAOC was highly homologous to AOC from other plant species. Phylogenetic analysis indicated that CsAOC was clustered in a closely related subgroup with AOC of Ipomoea nil. The full-length coding region of CsAOC was ligated with pET-32a and successfully expressed in E. coli BL21 (DE3), and purified. Real-time quantitative PCR analysis revealed that methyl jasmonate (MeJA) treatment on its own potently enhanced its expression over the control (healthy leaves), suggesting feedback activation of CsAOC. The expression under salicylic acid (SA) and wounding treatments was up-regulated. The mRNA expression of CsAOC could be induced by tea geometrids and tea green leafhoppers.

scholarly journals Biodegradation of Chloroxylenol by Cunninghamella elegans IM 1785/21GP and Trametes versicolor IM 373: Insight into Ecotoxicity and Metabolic Pathways

2021 ◽  
Vol 22 (9) ◽  
pp. 4360
Author(s):  
Marta Nowak ◽  
Katarzyna Zawadzka ◽  
Janusz Szemraj ◽  
Aleksandra Góralczyk-Bińkowska ◽  
Katarzyna Lisowska
Keyword(s):  
Trametes Versicolor ◽  
Significant Rise ◽  
Pcr Analysis ◽  
Cunninghamella Elegans ◽  
Oxidation Reactions ◽  
Cytochrome P450 Enzymes ◽  
C Elegans ◽  
Real Time Quantitative Pcr ◽  
Genes Expression ◽  
By Products

Chloroxylenol (PCMX) is applied as a preservative and disinfectant in personal care products, currently recommended for use to inactivate the SARS-CoV-2 virus. Its intensive application leads to the release of PCMX into the environment, which can have a harmful impact on aquatic and soil biotas. The aim of this study was to assess the mechanism of chloroxylenol biodegradation by the fungal strains Cunninghamella elegans IM 1785/21GP and Trametes versicolor IM 373, and investigate the ecotoxicity of emerging by-products. The residues of PCMX and formed metabolites were analysed using GC-MS. The elimination of PCMX in the cultures of tested microorganisms was above 70%. Five fungal by-products were detected for the first time. Identified intermediates were performed by dechlorination, hydroxylation, and oxidation reactions catalysed by cytochrome P450 enzymes and laccase. A real-time quantitative PCR analysis confirmed an increase in CYP450 genes expression in C. elegans cells. In the case of T. versicolor, spectrophotometric measurement of the oxidation of 2,20-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) showed a significant rise in laccase activity during PCMX elimination. Furthermore, with the use of bioindicators from different ecosystems (Daphtoxkit F and Phytotoxkit), it was revealed that the biodegradation process of PCMX had a detoxifying nature.

scholarly journals Molecular surveillance of shiga toxigenic Escherichia coli in selected beef abattoirs in Osun State Nigeria

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Femi Ayoade ◽  
Judith Oguzie ◽  
Philomena Eromon ◽  
Omolola E. Omotosho ◽  
Tosin Ogunbiyi ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Sequence Analysis ◽  
Block Design ◽  
Latex Agglutination ◽  
Pcr Analysis ◽  
Accession Number ◽  
E Coli ◽  
Representative Isolate ◽  
Randomized Block Design ◽  
Toxigenic Strains

AbstractShiga toxigenic strains of E. coli (STEC) known to be etiological agents for diarrhea were screened for their incidence/occurrence in selected abattoirs sources in Osogbo metropolis of Osun State, Nigeria using a randomized block design. Samples were plated directly on selective and differential media and E. coli isolates. Multiplex PCR analysis was used to screen for the presence of specific virulence factors. These were confirmed serologically as non-O157 STEC using latex agglutination serotyping kit. Sequence analysis of PCR products was performed on a representative isolate showing the highest combination of virulence genes using the 16S gene for identification purposes only. Results showed that the average cfu/cm2 was significantly lower in the samples collected at Sekona-2 slaughter slab compared with those collected at Al-maleek batch abattoir and Sekona-1 slaughter slab in ascending order at P = 0.03. Moreover, the average cfu/cm2E. coli in samples collected from butchering knife was significantly lower when compared with that of the workers’ hand (P = 0.047) and slaughtering floor (P = 0.047) but not with the slaughter table (P = 0.98) and effluent water from the abattoir house (P = 0.39). These data suggest that the abattoir type may not be as important in the prevalence and spread of STEC as the hygiene practices of the workers. Sequence analysis of a representative isolate showed 100% coverage and 96.46% percentage identity with Escherichia coli O113:H21 (GenBank Accession number: CP031892.1) strain from Canada. This sequence was subsequently submitted to GenBank with accession number MW463885. From evolutionary analyses, the strain from Nigeria, sequenced in this study, is evolutionarily distant when compared with the publicly available sequences from Nigeria. Although no case of E. coli O157 was found within the study area, percent occurrence of non-O157 STEC as high as 46.3% at some of the sampled sites is worrisome and requires regulatory interventions in ensuring hygienic practices at the abattoirs within the study area.

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