Identification of stage-specifically expressed genes of Trichinella spiralis by suppression subtractive hybridization

Parasitology ◽  
2007 ◽  
Vol 134 (10) ◽  
pp. 1443-1455 ◽  
Author(s):  
M. Y. LIU ◽  
X. L. WANG ◽  
B. Q. FU ◽  
C. Y. LI ◽  
X. P. WU ◽  
...  
Keyword(s):  
Suppression Subtractive Hybridization ◽  
Serine Proteases ◽  
Zinc Finger Protein ◽  
Trichinella Spiralis ◽  
Gene Families ◽  
Subtractive Hybridization ◽  
Cdna Clones ◽  
Dnase Ii ◽  
Finger Protein ◽  
Old Adult

SUMMARYNewborn larvae (NBL) and adult (Ad) stage-specifically expressed genes or members of gene families of Trichinella spiralis were identified by suppression subtractive hybridization (SSH)†. Six cDNA clones were identified as NBL stage-specific, including 1 member of the T. spiralis gene family encoding glutamic acid-rich proteins, 2 clones encoding novel serine proteases, 2 closely related clones encoding proteins that are members of a deoxyribonuclease II (DNase II)-like family and 1 clone with no similarity to known genes. Four stage-specific clones encoding homologues of retinoid X receptor, caveolin, C2H2 type zinc finger protein and a putative protein with no homology to known sequences were obtained from 3-day-old adult worms. One gene specifically up-regulated in the 5-day-old adult worms encoding a putative cuticle collagen was also identified.

Human proviral mRNAs down regulated in choriocarcinoma encode a zinc finger protein related to Krüppel

1990 ◽  
Vol 10 (8) ◽  
pp. 4401-4405 ◽  
Author(s):  
N Kato ◽  
K Shimotohno ◽  
D VanLeeuwen ◽  
M Cohen
Keyword(s):  
Amino Acid ◽  
Zinc Finger ◽  
Zinc Finger Protein ◽  
Genomic Sequences ◽  
Cdna Clones ◽  
Leader Region ◽  
Rna Transcripts ◽  
Finger Protein

RNA transcripts of the HERV-R (ERV3) human provirus that are abundant in placenta but absent in choriocarcinoma contain nonproviral genomic sequences at their 3' ends. We report here the isolation of cDNA clones of these genomic sequences. The transcripts encode a Krüppel-related zinc finger protein consisting of a unique leader region and more than 12 28-amino-acid finger motifs.

scholarly journals Proteomic Analysis ofTrichinella spiralisMuscle Larval Excretory-Secretory Proteins Recognized by Early Infection Sera

2013 ◽  
Vol 2013 ◽  
pp. 1-7 ◽  
Author(s):  
Li Wang ◽  
Zhong Quan Wang ◽  
Dan Dan Hu ◽  
Jing Cui
Keyword(s):  
Molecular Weight ◽  
Serine Proteases ◽  
Trichinella Spiralis ◽  
Early Stage ◽  
False Negative ◽  
Hypothetical Protein ◽  
Secretory Proteins ◽  
Dnase Ii ◽  
Negative Results ◽  
Protective Antibodies

Although the excretory-secretory (ES) proteins ofTrichinella spiralismuscle larvae are the most commonly used diagnostic antigens for trichinellosis, their main disadvantage is the false negative results during the early stage of infection and cross-reaction of their main components (43, 45, 49, and 53 kDa) with sera of patients with other helminthiasis. The aim of this study was to identify early specific diagnostic antigens inT. spiralisES proteins with 30–40 kDa. The ES proteins were analyzed by two-dimensional electrophoresis (2-DE), and a total of approximately 150 proteins spots were detected with isoelectric point (pI) varying from 4 to 7 and molecular weight from 14 to 66 kDa. When probed with sera from infected mice at 18 days postinfection, ten protein spots with molecular weight of 30–40 kDa were recognized and identified by MALDI-TOF/TOF-MS. All of ten spots were successfully identified and characterized to correlate with five different proteins, including two potential serine proteases, one antigen targeted by protective antibodies, one deoxyribonuclease (DNase) II, and one conserved hypothetical protein. These proteins might be the early specific diagnostic antigens for trichinellosis.

scholarly journals Identification of New Potential Regulators of the Medicago truncatula—Sinorhizobium meliloti Symbiosis Using a Large-Scale Suppression Subtractive Hybridization Approach

2007 ◽  
Vol 20 (3) ◽  
pp. 321-332 ◽  
Author(s):  
Laurence Godiard ◽  
Andreas Niebel ◽  
Fabienne Micheli ◽  
Jérôme Gouzy ◽  
Thomas Ott ◽  
...  
Keyword(s):  
Signal Transduction ◽  
Suppression Subtractive Hybridization ◽  
Medicago Truncatula ◽  
Large Scale ◽  
Sinorhizobium Meliloti ◽  
Zinc Finger Protein ◽  
Gene Clusters ◽  
Subtractive Hybridization ◽  
Symbiotic Interaction ◽  
Genes Encoding

We set up a large-scale suppression subtractive hybridization (SSH) approach to identify Medicago truncatula genes differentially expressed at different stages of the symbiotic interaction with Sinorhizobium meliloti, with a particular interest for regulatory genes. We constructed 7 SSH libraries covering successive stages from Nod factor signal transduction to S. meliloti infection, nodule organogenesis, and functioning. Over 26,000 clones were differentially screened by two rounds of macroarray hybridizations. In all, 3,340 clones, corresponding to genes whose expression was potentially affected, were selected, sequenced, and ordered into 2,107 tentative gene clusters, including 767 MtS clusters corresponding to new M. truncatula genes. In total, 52 genes encoding potential regulatory proteins, including transcription factors (TFs) and other elements of signal transduction cascades, were identified. The expression pattern of some of them was analyzed by quantitative reverse-transcription polymerase chain reaction in wild-type and in Nod¯ M. truncatula mutants blocked before or after S. meliloti infection. Three genes, coding for TFs of the bHLH and WRKY families and a C2H2 zinc-finger protein, respectively, were found to be upregulated, following S. meliloti inoculation, in the infection-defective mutant lin, whereas the bHLH gene also was expressed in the root-hair-curling mutant hcl. The potential role of these genes in early symbiotic steps is discussed.

scholarly journals Two novel non-cationic defensin-like antimicrobial peptides from haemolymph of the female tick, Amblyomma hebraeum

2004 ◽  
Vol 379 (3) ◽  
pp. 681-685 ◽  
Author(s):  
Ren LAI ◽  
Lee O. LOMAS ◽  
Jan JONCZY ◽  
Philip C. TURNER ◽  
Huw H. REES
Keyword(s):  
Antimicrobial Peptides ◽  
Suppression Subtractive Hybridization ◽  
Subtractive Hybridization ◽  
Protein Profiling ◽  
Differentially Expressed ◽  
Cdna Clones ◽  
Hard Tick ◽  
Differentially Expressed Proteins ◽  
Amblyomma Hebraeum ◽  
Cdna Sequences

Two non-cationic defensin-like antimicrobial peptides, named Amblyomma defensin peptide 1 and Amblyomma defensin peptide 2, were identified from the hard tick, Amblyomma hebraeum, by a combination of suppression subtractive hybridization for differentially expressed genes and proteomics. cDNA clones encoding each of these two defensin-like antimicrobial peptides were isolated from the differentially expressed cDNA library of the tick synganglia (central nervous system). The preproproteins deduced from the cDNA sequences each have 92 amino acid residues. Amblyomma defensin peptide 2 was purified from the haemolymph of fed female ticks. The purified peptide displayed antibacterial activity against Gram-negative and Gram-positive bacteria. Amblyomma defensin peptide 1 was further identified by protein chip capture combined with SELDI-TOF (surface-enhanced laser desorption/ionization–time-of-flight) MS. By screening for differentially expressed proteins, it was found that the expression of Amblyomma defensin peptide 1 was upregulated during 4 days post-feeding. Our findings firstly provide two defensin-like antimicrobial peptides that are particularly novel in being anionic, together with corresponding cDNA sequences, in hard ticks, and prove that the combination of suppression subtractive hybridization and protein profiling is a powerful method to study differentially expressed proteins, especially for organisms without available genome sequence information.

scholarly journals Identification of Defense-Related Rice Genes by Suppression Subtractive Hybridization and Differential Screening

2001 ◽  
Vol 14 (5) ◽  
pp. 685-692 ◽  
Author(s):  
Lizhong Xiong ◽  
Min-Woo Lee ◽  
Min Qi ◽  
Yinong Yang
Keyword(s):  
Suppression Subtractive Hybridization ◽  
Zinc Finger Protein ◽  
De Novo ◽  
Subtractive Hybridization ◽  
Defense Responses ◽  
Rice Cultivar ◽  
Differential Screening ◽  
Mitogen Activated Protein Kinase ◽  
Japonica Rice ◽  
Bacterial Colony

Identification of host genes involved in defense responses is one of most critical steps leading to the elucidation of disease resistance mechanisms in plants. In this study, two different cloning strategies were employed to identify defense-related genes from a tropical japonica rice cultivar (Oryza sativa cv. Drew). With the use of bacterial colony arrays, differential screening of a blast fungus (Pyricularia grisea)-induced rice cDNA library led to the isolation of 22 distinct rice genes that are expressed differentially in response to blast infection. Sequence analysis indicates that most of them are full-length cDNAs encoding pathogenesis-related proteins or other relatively abundant proteins. In combination with treatments of cycloheximide plus jasmonic acid (JA) or benzothiadiazole (BTH) in rice seedlings, the polymerase chain reaction-based suppression subtractive hybridization also was conducted to search for immediate early (IE) defense-related genes whose transcription is independent of de novo protein synthesis. The initial screening of only 768 subtracted clones resulted in the identification of 34 distinct IE genes that are induced by JA, BTH, and/or blast infection. Database searches revealed that these IE genes encode putative mitogen-activated protein kinase, diacylglycerol kinase, zinc finger protein, RelA-SpoT protein, ankyrin-containing protein, ABC transporter, β-ketoacyl-CoA synthase, and other potential defense-signaling components. Further characterization of these novel IE genes will likely facilitate the elucidation of defense signal transduction in rice plants.

scholarly journals Human proviral mRNAs down regulated in choriocarcinoma encode a zinc finger protein related to Krüppel.

1990 ◽  
Vol 10 (8) ◽  
pp. 4401-4405 ◽  
Author(s):  
N Kato ◽  
K Shimotohno ◽  
D VanLeeuwen ◽  
M Cohen
Keyword(s):  
Amino Acid ◽  
Zinc Finger ◽  
Zinc Finger Protein ◽  
Genomic Sequences ◽  
Cdna Clones ◽  
Leader Region ◽  
Rna Transcripts ◽  
Finger Protein

RNA transcripts of the HERV-R (ERV3) human provirus that are abundant in placenta but absent in choriocarcinoma contain nonproviral genomic sequences at their 3' ends. We report here the isolation of cDNA clones of these genomic sequences. The transcripts encode a Krüppel-related zinc finger protein consisting of a unique leader region and more than 12 28-amino-acid finger motifs.

scholarly journals ZBP-89, a Krüppel-like zinc finger protein, inhibits epidermal growth factor induction of the gastrin promoter.

1996 ◽  
Vol 16 (12) ◽  
pp. 6644-6653 ◽  
Author(s):  
J L Merchant ◽  
G R Iyer ◽  
B R Taylor ◽  
J R Kitchen ◽  
E R Mortensen ◽  
...  
Keyword(s):  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Zinc Finger ◽  
Zinc Finger Protein ◽  
Cell Receptor ◽  
Cdna Clones ◽  
Expression Library ◽  
Response Element ◽  
Finger Protein ◽  
Epidermal Growth

We have shown previously that a GC-rich element (GGGGCGGGGTGGGGGG) conferring epidermal growth factor (EGF) responsiveness to the human gastrin promoter binds Sp1 and additional undefined complexes. A rat GH4 cell line expression library was screened by using a multimer of the gastrin EGF response element, and three overlapping cDNA clones were identified. The full-length rat cDNA encoded an 89-kDa zinc finger protein (ZBP-89) that was 89% identical to a 49-kDa human factor, ht(beta), that binds a GTGGG/CACCC element in T-cell receptor promoters. The conservation of amino acids between the zinc fingers indicates that ZBP-89 is a member of the C2H2 zinc finger family subclass typified by the Drosophila Krüppel protein. ZBP-89 is ubiquitously expressed in normal adult tissues. It binds specifically to the gastrin EGF response element and inhibits EGF induction of the gastrin promoter. Collectively, these results demonstrate that ZBP-89 functions as a repressor of basal and inducible expression of the gastrin gene.

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