Suppression of Transferrin Expression Enhances the Susceptibility of Plutella xylostella to Isaria cicadae

Transferrins (Trfs) are multifunctional proteins with key functions in iron transport. In the present study, a Trf (PxTrf) from Plutella xylostella was identified and characterized. The PxTrf consisted of a 2046-bp open reading frame, which encoded a 681 amino acid protein with a molecular weight of 73.43 kDa and had an isoelectric point of 7.18. Only a single iron domain was predicted in the N-lobe of PxTrf. Although PxTrf was expressed ubiquitously, the highest levels of expression were observed in the fourth instar larvae. PxTrf transcript level was highest in fat bodies among various tissues. The PxTrf transcript levels increased significantly after the stimulation of pathogens. A decrease in PxTrf expression via RNA interference enhanced the susceptibility of P. xylostella to the Isaria cicadae fungus and inhibited hemocyte nodulation in response to the fungal challenge. In addition, a considerable increase in the pupation rate was observed in larvae treated with double-stranded PxTrf (dsPxTrf). Overall, according to the results, PxTrf may participate in P. xylostella immunity against fungal infection and insect development.

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The bglA Gene of Aspergillus kawachii Encodes Both Extracellular and Cell Wall-Bound β-Glucosidases

Applied and Environmental Microbiology ◽

10.1128/aem.65.12.5546-5553.1999 ◽

1999 ◽

Vol 65 (12) ◽

pp. 5546-5553 ◽

Cited By ~ 37

Author(s):

Kazuhiro Iwashita ◽

Tatsuya Nagahara ◽

Hitoshi Kimura ◽

Makoto Takano ◽

Hitoshi Shimoi ◽

...

Keyword(s):

Cell Wall ◽

Amino Acid ◽

Amino Acid Sequence ◽

Amino Acid Sequences ◽

Enzyme Assays ◽

Partial Amino Acid Sequence ◽

Reading Frame ◽

Acid Protein ◽

Aspergillus Kawachii ◽

Amino Acid Protein

ABSTRACT We cloned the genomic DNA and cDNA of bglA, which encodes β-glucosidase in Aspergillus kawachii, based on a partial amino acid sequence of purified cell wall-bound β-glucosidase CB-1. The nucleotide sequence of the cloned bglA gene revealed a 2,933-bp open reading frame with six introns that encodes an 860-amino-acid protein. Based on the deduced amino acid sequence, we concluded that the bglA gene encodes cell wall-bound β-glucosidase CB-1. The amino acid sequence exhibited high levels of homology with the amino acid sequences of fungal β-glucosidases classified in subfamily B. We expressed the bglA cDNA inSaccharomyces cerevisiae and detected the recombinant β-glucosidase in the periplasm fraction of the recombinant yeast.A. kawachii can produce two extracellular β-glucosidases (EX-1 and EX-2) in addition to the cell wall-bound β-glucosidase.A. kawachii in which the bglA gene was disrupted produced none of the three β-glucosidases, as determined by enzyme assays and a Western blot analysis. Thus, we concluded that thebglA gene encodes both extracellular and cell wall-bound β-glucosidases in A. kawachii.

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Molecular cloning, characterization, and expression analysis of a novel sema-2a homologue in Polyrhachis vicina (Hymenoptera: Formicidae)

The Canadian Entomologist ◽

10.4039/n09-046 ◽

2010 ◽

Vol 142 (1) ◽

pp. 1-13 ◽

Cited By ~ 2

Author(s):

Jing Luo ◽

Geng-Si Xi ◽

Shu-Min Lü ◽

Ke Li ◽

Qing Li

Keyword(s):

Untranslated Region ◽

Axonal Guidance ◽

Mrna Levels ◽

Chain Reaction ◽

Base Pairs ◽

Reading Frame ◽

Acid Protein ◽

Polymerase Chain ◽

Polyrhachis Vicina ◽

Amino Acid Protein

AbstractThe semaphorin gene family plays important roles in axonal guidance in vertebrates and invertebrates. Semaphorin 2a, a member of the semaphorin family, belongs to class 2, which is found only in invertebrates. In our study, semaphorin 2a was cloned from the ant Polyrhachis vicina Roger. The full length of P. vicina semaphorin 2a (Pv-sema-2a) is 2763 base pairs (bp) and it contains a 5′-untranslated region (UTR) 92 bp long and a 3′-UTR 521 bp long. The open reading frame of Pv-sema-2a encodes a 716-amino-acid protein with a predicted molecular mass of 81.1 kilodaltons. Real-time quantitative reverse-transcription – polymerase chain reaction indicated that Pv-sema-2a mRNA is differentially expressed during P. vicina development, in the whole bodies as well as the heads of different castes. The high mRNA levels in embryos and pupae suggest that Pv-sema-2a plays an important role in ant development.

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New Carbenicillin-Hydrolyzing β-Lactamase (CARB-7) from Vibrio cholerae Non-O1, Non-O139 Strains Encoded by the VCR Region of the V. cholerae Genome

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.46.7.2162-2168.2002 ◽

2002 ◽

Vol 46 (7) ◽

pp. 2162-2168 ◽

Cited By ~ 30

Author(s):

Roberto Melano ◽

Alejandro Petroni ◽

Alicia Garutti ◽

Héctor Alex Saka ◽

Laura Mange ◽

...

Keyword(s):

Nucleotide Sequence ◽

Vibrio Cholerae ◽

Gene Families ◽

Chromosome 2 ◽

Reading Frame ◽

West Region ◽

Acid Protein ◽

Lactamase Gene ◽

Flanking Regions ◽

Amino Acid Protein

ABSTRACT In a previous study, an analysis of 77 ampicillin-nonsusceptible (resistant plus intermediate categories) strains of Vibrio cholerae non-O1, non-O139, isolated from aquatic environment and diarrheal stool, showed that all of them produced a β-lactamase with a pI of 5.4. Hybridization or amplification by PCR with a probe for bla TEM or primers for bla CARB gene families was negative. In this work, an environmental ampicillin-resistant strain from this sample, ME11762, isolated from a waterway in the west region of Argentina, was studied. The nucleotide sequence of the structural gene of the β-lactamase was determined by bidirectional sequencing of a Sau3AI fragment belonging to this isolate. The gene encodes a new 288-amino-acid protein, designated CARB-7, that shares 88.5% homology with the CARB-6 enzyme; an overall 83.2% homology with PSE-4, PSE-1, CARB-3, and the Proteus mirabilis N29 enzymes; and 79% homology with CARB-4 enzyme. The gene for this β-lactamase could not be transferred to Escherichia coli by conjugation. The nucleotide sequence of the flanking regions of the bla CARB-7 gene showed the occurrence of three 123-bp V. cholerae repeated sequences, all of which were found outside the predicted open reading frame. The upstream fragment of the bla CARB-7 gene shared 93% identity with a locus situated inside V. cholerae's chromosome 2. These results strongly suggest the chromosomal location of the bla CARB-7 gene, making this the first communication of a β-lactamase gene located on the VCR island of the V. cholerae genome.

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Cloning and characterization of a 2,3-oxidosqualene cyclase from Eleutherococcus trifoliatus

Holzforschung ◽

10.1515/hf-2012-0120 ◽

2013 ◽

Vol 67 (4) ◽

pp. 463-471 ◽

Cited By ~ 1

Author(s):

Li-Ting Ma ◽

Sheng-Yang Wang ◽

Yen-Hsueh Tseng ◽

Yi-Ru Lee ◽

Fang-Hua Chu

Keyword(s):

Physiological Role ◽

Active Site Residues ◽

Reading Frame ◽

Oxidosqualene Cyclases ◽

Oxidosqualene Cyclase ◽

Acid Protein ◽

Leaf Tissues ◽

New Perspective ◽

Amino Acid Protein

Abstract The 2,3-oxidosqualene cyclases (OSCs) are a family of enzymes that have an important role in plant triterpene biosynthesis. In this study, an OSC gene designed EtLUS from Eleutherococcus trifoliatus has been cloned. EtLUS includes a 2292-bp open reading frame and encodes a 763-amino acid protein. EtLUS has an MLCYCR motif, which is conserved in lupeol synthases. Comparison of active-site residues and gene expression in yeast showed that EtLUS synthesizes lupeol. However, EtLUS has the highest sequence identity with β-amyrin synthases from Araliaceae rather than lupeol synthases, adding new perspective to the evolution of the OSCs of Araliaceae. Furthermore, EtLUS is upregulated in leaf tissues under methyl jasmonate treatment, which can be interpreted that lupeol and its derivatives play an ecological and physiological role in plant defense against pathogens and insect herbivores.

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Identification and characterization of a cDNA encoding mouse CAP: a homolog of the yeast adenylyl cyclase associated protein

Journal of Cell Science ◽

10.1242/jcs.105.3.777 ◽

1993 ◽

Vol 105 (3) ◽

pp. 777-785 ◽

Cited By ~ 4

Author(s):

A.B. Vojtek ◽

J.A. Cooper

Keyword(s):

Adenylyl Cyclase ◽

Leading Edge ◽

Northern Analysis ◽

Reading Frame ◽

Terminal Domain ◽

Carboxy Terminal Domain ◽

Acid Protein ◽

Carboxy Terminal ◽

Amino Acid Protein

CAP, an adenylyl cyclase associated protein, is present in Saccharomyces cerevisiae and Schizosaccharomyces pombe. In both organisms, CAP is bifunctional: the N-terminal domain binds to adenylyl cyclase, thereby enabling adenylyl cyclase to respond appropriately to upstream regulatory signals, such as RAS in S. cerevisiae; the C-terminal domain is required for cellular morphogenesis. Here, we describe the isolation of a cDNA encoding a CAP homolog from a higher eukaryote. The mouse CAP cDNA contains an open reading frame capable of encoding a 474 amino acid protein. The protein encoded by the mouse CAP cDNA shows extensive homology to the yeast CAP proteins, particularly in the central poly-proline rich region and in the C-terminal domain. By northern analysis, the CAP message appears to be ubiquitous, but not uniform. By indirect immunofluorescence, ectopically expressed mouse CAP protein is found in the cytoplasm of fibroblasts and, in migrating cells, at the leading edge. Expression of the mouse CAP cDNA in S. cerevisiae complements defects associated with loss of the yeast CAP carboxy-terminal domain. Hence, the function of the CAP carboxy-terminal domain has been conserved from yeast to mouse.

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Molecular Cloning and Characterization of a cDNA Encoding Sedoheptulose-1,7-bisphosphatase from Mulberry (Morus alba var. multicaulis)

Silvae Genetica ◽

10.1515/sg-2008-0023 ◽

2008 ◽

Vol 57 (1-6) ◽

pp. 152-157 ◽

Cited By ~ 3

Author(s):

X. Ji ◽

Y. Gai ◽

J. Ma ◽

C. Zheng ◽

Z. Mu

Keyword(s):

Morus Alba ◽

Evolutionary Analysis ◽

Comparison Analysis ◽

Reading Frame ◽

Acid Protein ◽

Fructose 1,6 Bisphosphatase ◽

Rapid Amplification ◽

Molecular Evolutionary Analysis ◽

Amino Acid Protein

Abstract A full-length cDNA encoding sedoheptulose-1,7-bisphosphatase (SBPase; EC 3.1.3.37) was cloned from mulberry (Morus alba var. multicaulis) by rapid amplification of cDNA ends (RACE). The cDNA consisted of 1,527 nucleotides with an open reading frame (ORF) of 1,179 nucleotides encoding a 393 amino acid protein of approximately 42.6 kDa. Sequence comparison analysis showed that mulberry SBPase (MSBPase) had high homology to other plant counterparts. Phylogenetic and molecular evolutionary analysis revealed that MSBPase fell into plant SBPase group. Moreover, SBPase and fructose-1,6-bisphosphatase (FBPase; EC 3.1.3.11) shared 28-32% identical residues, suggesting that the two enzymes originated from the same evolution branch. Molecular modeling indicated that each subunit of MSBPase was composed of α-helices and β-sheets joined by turns and loops, and folded into a structure of hexahedron shape which was very similar to FBPase.

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NOP3 is an essential yeast protein which is required for pre-rRNA processing.

The Journal of Cell Biology ◽

10.1083/jcb.119.4.737 ◽

1992 ◽

Vol 119 (4) ◽

pp. 737-747 ◽

Cited By ~ 57

Author(s):

I D Russell ◽

D Tollervey

Keyword(s):

Northern Analysis ◽

Rna Recognition Motif ◽

Rna Recognition ◽

Rrna Processing ◽

Reading Frame ◽

Repeat Units ◽

Nucleolar Proteins ◽

Acid Protein ◽

Sds Page ◽

Amino Acid Protein

The four nucleolar proteins NOP1, SSB1, GAR1, and NSR1 of Saccharomyces cerevisiae share a repetitive domain composed of repeat units rich in glycine and arginine (GAR domain). We have cloned and sequenced a fifth member of this family, NOP3, and shown it to be essential for cell viability. The NOP3 open reading frame encodes a 415 amino acid protein with a predicted molecular mass of 45 kD, containing a GAR domain and an RNA recognition motif. NOP3-specific antibodies recognize a 60-kD protein by SDS-PAGE and decorate the nucleolus and the surrounding nucleoplasm. A conditional lethal mutation, GAL::nop3, was constructed; growth of the mutant strain in glucose medium represses NOP3 expression. In cells depleted of NOP3, production of cytoplasmic ribosomes is impaired. Northern analysis and pulse-chase labeling indicate that pre-rRNA processing is inhibited at the late steps, in which 27SB pre-rRNA is cleaved to 25S rRNA and 20S pre-rRNA to 18S rRNA.

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Characterization of an Endo-β-1,6-Galactanase from Streptomyces avermitilis NBRC14893

Applied and Environmental Microbiology ◽

10.1128/aem.01733-07 ◽

2008 ◽

Vol 74 (8) ◽

pp. 2379-2383 ◽

Cited By ~ 16

Author(s):

Hitomi Ichinose ◽

Toshihisa Kotake ◽

Yoichi Tsumuraya ◽

Satoshi Kaneko

Keyword(s):

Signal Sequence ◽

Recombinant Enzyme ◽

Streptomyces Avermitilis ◽

Glycoside Hydrolase Family ◽

Reading Frame ◽

Secretion Signal ◽

Acid Protein ◽

Hydrolysis Of ◽

Amino Acid Protein

ABSTRACT The putative endo-β-1,6-galactanase gene from Streptomyces avermitilis was cloned and expressed in Escherichia coli, and the enzymatic properties of the recombinant enzyme were characterized. The gene consisted of a 1,476-bp open reading frame and encoded a 491-amino-acid protein, comprising an N-terminal secretion signal sequence and glycoside hydrolase family 5 catalytic module. The recombinant enzyme, Sa1,6Gal5A, catalyzed the hydrolysis of β-1,6-linked galactosyl linkages of oligosaccharides and polysaccharides. The enzyme produced galactose and a range of β-1,6-linked galacto-oligosaccharides, predominantly β-1,6-galactobiose, from β-1,6-galactan chains. There was a synergistic effect between the enzyme and Sa1,3Gal43A in degrading tomato arabinogalactan proteins. These results suggest that Sa1,6Gal5A is the first identified endo-β-1,6-galactanase from a prokaryote.

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Cloning and expression of a novel UDP-GlcNAc:α-d-mannoside β1,2-N-acetylglucosaminyltransferase homologous to UDP-GlcNAc:α-3-d-mannoside β1,2-N-acetylglucosaminyltransferase I

Biochemical Journal ◽

10.1042/bj3610153 ◽

2001 ◽

Vol 361 (1) ◽

pp. 153-162 ◽

Cited By ~ 20

Author(s):

Wenli ZHANG ◽

Doron BETEL ◽

Harry SCHACHTER

Keyword(s):

Northern Blot Analysis ◽

Human Gene ◽

Cloning And Expression ◽

Chromosome 1 ◽

Tblastn Search ◽

Reading Frame ◽

Acid Protein ◽

Human And Mouse ◽

Amino Acid Protein

A TBLASTN search with human UDP-GlcNAc:α-3-d-mannoside β-1,2-N-acetylglucosaminyltransferase I (GnT I; EC 2.4.1.101) as a probe identified human and mouse Unigenes encoding a protein similar to human GnT I (34% identity over 340 amino acids). The recombinant protein converted Man(α1–6)[Man(α1–3)]Man(β1-)O-octyl to Man(α1–6)[GlcNAc(β1–2)Man(α1–3)]Man(β1-)O-octyl, the reaction catalysed by GnT I. The enzyme also added GlcNAc to Man(α1–6)[GlcNAc(β1–2)Man(α1–3)]Man(β1-)O-octyl (the substrate for β-1,2-N-acetylglucosaminyltransferase II), Man(α1-)O-benzyl [with Km values of ≈ 0.3 and > 30mM for UDP-GlcNAc and Man(α1-)O-benzyl respectively] and the glycopeptide CYA[Man(α1-)O-T]AV (Km ∼ 12mM). The product formed with Man(α1-)O-benzyl was identified as GlcNAc(β1–2)Man(α1-)O-benzyl by proton NMR spectroscopy. The enzyme was named UDP-GlcNAc:α-d-mannoside β-1,2-N-acetylglucosaminyltransferase I.2 (GnT I.2). The human gene mapped to chromosome 1. Northern-blot analysis showed a 3.3kb message with a wide tissue distribution. The cDNA has a 1980bp open reading frame encoding a 660 amino acid protein with a type-2 domain structure typical of glycosyltransferases. Man(β1-)O-octyl, Man(β1-)O-p-nitrophenyl and GlcNAc(β1–2)Man(α1–6)[GlcNAc(β1–2)Man(α1–3)]Man(β1–4)GlcNAc(β1–4)GlcNAc(β1-)O-Asn were not acceptors, indicating that GnT I.2 is specific for α-linked terminal Man and does not have N-acetylglucosaminyltransferase III, IV, V, VII or VIII activities. CYA[Man(α1-)O-T]AV was between three and seven times more effective as an acceptor than the other substrates, suggesting that GnT I.2 may be responsible for the synthesis of the GlcNAc(β1–2)Man(α1-)O-Ser/Thr moiety on α-dystroglycan and other O-mannosylated proteins.

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Molecular Cloning and Biochemical Characterization of a Recombinant Sterol 3-O-Glucosyltransferase fromGymnema sylvestreR.Br. Catalyzing Biosynthesis of Steryl Glucosides

BioMed Research International ◽

10.1155/2014/934351 ◽

2014 ◽

Vol 2014 ◽

pp. 1-14 ◽

Cited By ~ 10

Author(s):

Pragya Tiwari ◽

Rajender Singh Sangwan ◽

Asha ◽

B. N. Mishra ◽

Farzana Sabir ◽

...

Keyword(s):

Biochemical Characterization ◽

Present Report ◽

Hydroxyl Group ◽

Biological Origin ◽

Reading Frame ◽

Steryl Glucosides ◽

Acid Protein ◽

Phytochemical Profiles ◽

Amino Acid Protein

Gymnema sylvestreR.Br., a pharmacologically important herb vernacularly called Gur-Mar (sugar eliminator), is widely known for its antidiabetic action. This property of the herb has been attributed to the presence of bioactive triterpene glycosides. Although some information regarding pharmacology and phytochemical profiles of the plant are available, no attempts have been made so far to decipher the biosynthetic pathway and key enzymes involved in biosynthesis of steryl glucosides. The present report deals with the identification and catalytic characterization of a glucosyltransferase, catalyzing biosynthesis of steryl glycosides. The full length cDNA (2572 bp) contained an open reading frame of 2106 nucleotides that encoded a 701 amino acid protein, falling into GT-B subfamily of glycosyltransferases. The GsSGT was expressed inEscherichia coliand biochemical characterization of the recombinant enzyme suggested its key role in the biosynthesis of steryl glucosides with catalytic preference for C-3 hydroxyl group of sterols. To our knowledge, this pertains to be the first report on cloning and biochemical characterization of a sterol metabolism gene fromG. sylvestreR.Br. catalyzing glucosylation of a variety of sterols of biological origin from diverse organisms such as bacteria, fungi, and plants.

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