Control of polygalacturonase synthesis inFusarium oxyspotumf.sp.radicis lycopersici

1997 ◽  
Vol 43 (11) ◽  
pp. 1084-1090 ◽  
Author(s):  
Belén Patiño ◽  
Martha Lucía Posada ◽  
Covadonga Vázquez ◽  
María Teresa González-Jaén ◽  
Álvaro Martínez del Pozo
Keyword(s):  
Fusarium Oxysporum ◽  
Blot Analysis ◽  
Genomic Dna ◽  
Southern Blot Analysis ◽  
Northern Blot Analysis ◽  
Single Copy ◽  
Transcriptional Level ◽  
High Similarity ◽  
Apple Pectin ◽  
Sds Page

Genetic control of polygalacturonase (PG) activity from Fusarium oxysporum f.sp. radicis lycopersici was analyzed on pectin and glucose cultures. One exopolygalacturonase from F. oxysporum f.sp. radicis lycopersici was strongly induced, in stationary culture, when the fungus was grown on apple pectin, while on glucose no extracellular PG activity could be detected. Although SDS–PAGE detected the presence of a putative PG band (66 kDa) in both conditions, specific antibodies obtained against the purified PG only detected it in PG-inducing conditions, that is to say, when apple pectin was used as the carbon source. Northern blot analysis of RNA of two isolates of F. oxysporum f.sp. radicis lycopersici (r6and r2) confirmed that this regulation of PG synthesis was exerted at the transcriptional level. Only one single mRNA species of around 1400 nucleotides was detected on the cultures containing pectin and was absent in glucose-grown cultures. Southern blot analysis of genomic DNA indicated that pg gene seems to be present in a single copy in the genomes of F. oxysporum f.sp. radicis lycopersici r6and r2and Fusarium oxysporum f.sp. lycopersici, showing similar hybridization patterns in all species. The partial sequence of this pg gene from F. oxysporum f.sp. radicis lycopersici r6, which is also reported, showed high similarity to diverse PGs already reported. Exopolygalacturonase of F. oxysporum f.sp. radicis lycopersici r6is heavily glycosylated; its deglycosylated form had a molecular mass of 50 kDa.Key words: polygalacturonase, Fusarium oxysporum f.sp. radicis lycopersici, regulation.

scholarly journals Tissue distribution of rat glutathione transferase subunit 7, a hepatoma marker

1986 ◽  
Vol 240 (3) ◽  
pp. 885-889 ◽  
Author(s):  
S E Pemble ◽  
J B Taylor ◽  
B Ketterer
Keyword(s):  
Northern Blot ◽  
Blot Analysis ◽  
Genomic Dna ◽  
Southern Blot Analysis ◽  
Northern Blot Analysis ◽  
Glutathione Transferase ◽  
Rat Tissues ◽  
Polyadenylated Rna ◽  
Normal Rat ◽  
Rat Hepatoma

Polyadenylated RNA isolated from NN-dimethyl-4-aminoazobenzene-induced rat hepatoma was used to prepare a cDNA library in lambda gt10. Full-length clones complementary to mRNA coding for glutathione transferase subunit 7 were isolated and one of these clones (pGSTr7) was fully characterized. In Northern blot analysis, mRNA hybridizing to 32P-labelled pGSTr7 was found in poly(A)-containing RNA isolated from seven normal rat tissues but not from testis and liver. A similar hybridizing mRNA species was also detected in human placental mRNA. The same probe, used in a Southern blot analysis of genomic DNA, suggests the presence of a multigene family in the rat.

scholarly journals Synthesis and glycosylation of CD52, the major 'maturation-associated' antigen of rat spermatozoa, in the cauda epididymidis

Reproduction ◽  
2001 ◽  
pp. 435-446 ◽  
Author(s):  
P Derr ◽  
CH Yeung ◽  
TG Cooper ◽  
C Kirchhoff
Keyword(s):  
Blot Analysis ◽  
Cdna Sequence ◽  
Northern Blot Analysis ◽  
Lectin Binding ◽  
Transcriptional Level ◽  
Lectin Blot ◽  
Molecular Masses ◽  
Rat Epididymis ◽  
Sperm Membrane ◽  
Cauda Epididymidis

A western and lectin blot analysis was performed of the major 'maturation-associated' antigen of rat spermatozoa, which is the rat counterpart of human CD52. In the absence of a suitable antibody, direct study of this approximately 26 kDa antigen, named previously SMemG, had been difficult. In the present study, these problems were overcome by raising a polyclonal antibody against a chemosynthetic peptide predicted from the cDNA sequence of the antigen. The antibody bound to a glycoprotein of rat cauda epididymidal tissue and spermatozoa, this glycoprotein was cleaved by phosphatidylinositol-specific phospholipase C and, after deglycosylation, was reduced to approximately 6 kDa. Northern blot analysis confirmed that the CD52 mRNA was transcribed only post-testicularly, and antibody binding to testicular and sperm proteins of different molecular masses was shown to be nonspecific. Flow cytometry also indicated that the antigen was inserted into the sperm membrane during epididymal transit. Moreover, despite the presence of CD52 mRNA in all parts of the rat epididymis, only the 'long' mRNA molecules of the cauda region were efficiently translated and the antigen glycosylated, indicating that expression of rat CD52 is regulated on a post-transcriptional level. Lectin binding and deglycosylation studies supported the contention that there is extensive mucin-type O-glycosylation of rat CD52. In rats, there was no indication of complex N-linked carbohydrates similar to those described for human CD52.

scholarly journals bcr Rearrangement without juxtaposition of c-abl in chronic myelocytic leukemia.

1985 ◽  
Vol 162 (6) ◽  
pp. 2175-2179 ◽  
Author(s):  
C R Bartram
Keyword(s):  
Blot Analysis ◽  
Southern Blot Analysis ◽  
Gene Rearrangement ◽  
Northern Blot Analysis ◽  
Philadelphia Chromosome ◽  
Leukemic Cells ◽  
Chronic Myelocytic Leukemia ◽  
Myelocytic Leukemia ◽  
Genomic Recombination

Southern blot analysis detected a bcr gene rearrangement within leukemic cells of a Philadelphia chromosome-negative chronic myelocytic leukemia (CML) patient that led to transcription of a novel 7.3 kb bcr RNA species. Participation of the c-abl oncogene in this genomic recombination could be ruled out by in situ hybridization studies and Northern blot analysis.

scholarly journals Expression of transcobalamin II mRNA in human tissues and cultured fibroblasts from normal and transcobalamin II-deficient patients

1994 ◽  
Vol 301 (2) ◽  
pp. 585-590 ◽  
Author(s):  
N Li ◽  
S Seetharam ◽  
D S Rosenblatt ◽  
B Seetharam
Keyword(s):  
Southern Blot ◽  
Blot Analysis ◽  
Genomic Dna ◽  
Southern Blot Analysis ◽  
Sequence Similarity ◽  
Ribonuclease Protection Assay ◽  
Rat Tissues ◽  
Cultured Fibroblasts ◽  
Ribonuclease Protection ◽  
Transcobalamin Ii

Transcobalamin II (TCII) is an important plasma transporter of cobalamin (Cbl; vitamin B12). In the present study, TCII gene expression in human and rat tissues and in the fibroblasts of patients with TCII deficiency was investigated. Northern-blot analyses revealed expression of TCII mRNA in many human and rat tissues. In humans, this was 14-fold higher in the kidney than in liver, whereas in the rat the levels of expression were similar in the kidney and liver. Southern-blot analysis of genomic DNA from several species revealed sequence similarity in TCII across species. Metabolic labelling and ribonuclease protection assay revealed a 43 kDa TCII protein and a fully protected TCII mRNA band in normal fibroblasts but not in fibroblasts from three TCII-deficient patients. Southern-blot analysis of genomic DNA from all these fibroblasts revealed identical restriction patterns on BamHI, HindIII, KpnI, MspI and EcoRI digestion. On the basis of these results, we suggest that TCII is expressed in multiple tissues, and its level of expression in tissues varies within the same and across species. Furthermore, the TCII deficiency characterized in this study is due to the absence of TCII protein which in turn is due to the absence or extremely low levels of its mRNA and not to detectable gross alterations in the gene structure.

ADictyostelium discoideumgene, which is highly related tomo15fromXenopus, is expressed during growth but not during development

1995 ◽  
Vol 73 (1-2) ◽  
pp. 51-58 ◽  
Author(s):  
Christine Michaelis ◽  
Qian Luo ◽  
Gerald Weeks
Keyword(s):  
Blot Analysis ◽  
Northern Blot Analysis ◽  
Single Copy ◽  
Cellular Slime Mold ◽  
Cell Type ◽  
Cdc2 Kinase ◽  
Related Family ◽  
Copy Gene ◽  
Slime Mold Dictyostelium Discoideum ◽  
Cellular Slime

We have isolated a cDNA from the cellular slime mold Dictyostelium discoideum encoding a protein that is 52% identical to the Xenopus Mo15 kinase and highly related to the equivalent proteins from human (52% identity), rice (52.7% identity), and yeast (47.6% identity). Mo15 is responsible for the activation of Cdc2 kinase and is itself a member of the large Cdc2-related family of protein kinases. The Dictyostelium protein is more related to the Xenopus Mo15 protein than it is to either the Dictyostelium Cdc2 or Crp proteins. Southern blot analysis of genomic V12-M2 DNA indicated that mo15 is present as a single copy gene that cross hybridizes with cdc2 at low stringency. Northern blot analysis of RNA from different stages of Dictyostelium development showed that mo15 is only expressed during vegetative cell growth.Key words: cell cycle, differentiation, cell type determination, Cdc2 kinase, Dictyostelium, Mo15 protein.

scholarly journals Rearrangement and overexpression of the BCL-1/PRAD-1 gene in intermediate lymphocytic lymphomas and in t(11q13)-bearing leukemias

Blood ◽  
1993 ◽  
Vol 81 (11) ◽  
pp. 3063-3067 ◽  
Author(s):  
R Rimokh ◽  
F Berger ◽  
G Delsol ◽  
C Charrin ◽  
MF Bertheas ◽  
...  
Keyword(s):  
Northern Blot ◽  
Blot Analysis ◽  
Southern Blot Analysis ◽  
Northern Blot Analysis ◽  
Lymphoid Tissues ◽  
Over Expression ◽  
Lymphoid Neoplasia ◽  
Flanking Region ◽  
Variant Translocation

Abstract The t(11;14)(q13;q32) translocation and its molecular counterpart, BCL- 1 rearrangement, are consistent features of intermediate lymphocytic lymphoma (ILL). Rearrangement is thought to deregulate the nearby PRAD- 1/BCL-1 proto-oncogene that is a newly identified member of the cyclin family. To characterize further the association between rearrangement of chromosome 11q13 and over-expression of BCL-1. Southern blot analysis was performed in 33 cases of ILL, 5 cases of t(11;14)- associated leukemias, and 1 case of leukemia carrying a variant translocation t(11;19)(q13;q13) using three separate BCL-1 locus probes. When RNA was available, BCL-1 expression was assessed by Northern blot analysis. DNA from 19 of 33 ILL (57%) showed BCL-1 rearrangement, 16 involving the major translocation cluster (MTC) region and 3 involving a new breakpoint cluster located in the 5' flanking region of the BCL-1 gene. DNA from 3 of 6 t(11q13)-associated leukemias demonstrated a rearrangement involving the MTC. Northern blot analysis showed that BCL-1 was overexpressed in 14 of 15 ILL and in all leukemias analyzed (included the t(11;19) leukemia) relative to normal and malignant lymphoid tissues. These results constitute additional elements in favor of the role of BCL-1 in lymphoid neoplasia and allow us to speculate about its mechanisms of activation.

scholarly journals Treatment of genomic DNA with T4 DNA ligase improves Southern blot analysis

1988 ◽  
Vol 16 (21) ◽  
pp. 10387-10387 ◽  
Author(s):  
Jørn Erland Koch ◽  
Steen Kølvraa ◽  
Morten Corneliusen ◽  
Kirsten Brunn Petersen ◽  
Niels Gregersen
Keyword(s):  
Southern Blot ◽  
Blot Analysis ◽  
Genomic Dna ◽  
Southern Blot Analysis ◽  
Dna Ligase ◽  
T4 Dna Ligase

Analysis of NLS and rRNA binding motifs in the L25 ribosomal protein from Leishmania (Viannia) braziliensis: investigation of its diagnostic capabilities

Parasitology ◽  
2002 ◽  
Vol 125 (1) ◽  
pp. 51-57 ◽  
Author(s):  
A. C. GONZÁLEZ ◽  
E. MARTÍNEZ ◽  
E. CARMELO ◽  
J. E. PIÑERO ◽  
V. ALONSO ◽  
...  
Keyword(s):  
Ribosomal Protein ◽  
Blot Analysis ◽  
Southern Blot Analysis ◽  
Northern Blot Analysis ◽  
Logarithmic Phase ◽  
Leishmania Braziliensis ◽  
Binding Motifs ◽  
Binding Domains ◽  
Antigenic Properties ◽  
Diagnostic Capabilities

A cDNA clone codifying ribosomal protein L25 was isolated from a Leishmania braziliensis cDNA gene library. The alignment of the amino acid sequence deduced from this gene with other proteins revealed that this protein is related to the L23/25 ribosomal protein family. This is so because this protein shows, in its C-terminal end, the rRNA binding domains characteristic of these proteins and at the N-terminal end the NLS sequence necessary for its entry into the nucleus. Southern blot analysis showed 2 copies of gene L25 per genome arranged in tandem position and pointing in the same direction. Northern blot analysis showed that this gene is transcribed in 2 mRNAs when parasite promastigotes are in the logarithmic phase. In order to analyse the antigenic properties of L. braziliensis RPL25, it was purified as a recombinant protein and ELISA-tested against cutaneous, mucocutaneous and Chagasic sera. The results indicate that the recombinant RPL25 from L. braziliensis presents a non-specific reaction that disqualifies it for the diagnosis of cutaneous leishmaniasis. In contrast, some of the synthetic peptides derived from its sequence may serve as promising tools for the diagnosis of this disease.

scholarly journals Establishment of cell lines from the wasp Hyposoter didymator (Hym., Ichneumonidae) containing the symbiotic polydnavirus H. didymator ichnovirus

2004 ◽  
Vol 85 (4) ◽  
pp. 863-868 ◽  
Author(s):  
Janick Rocher ◽  
Marc Ravallec ◽  
Patrick Barry ◽  
Anne-Nathalie Volkoff ◽  
Dominique Ray ◽  
...  
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Blot Analysis ◽  
Southern Blot Analysis ◽  
Northern Blot Analysis ◽  
High Molecular Mass ◽  
Dna Virus ◽  
Viral Dna ◽  
Hyposoter Didymator ◽  
Gene Expression Levels

Cell lines derived from polydnavirus-associated wasps should constitute a valuable tool for investigations of polydnavirus replication, but none is yet available. In this work, we describe the first cell lines, named Hd-AA, -AD, -BBA and -K, to have been established from the ichneumonid wasp Hyposoter didymator, associated with the polydnavirus H. didymator ichnovirus (HdIV). Southern blot analysis indicated that the viral DNA was present in all four cell lines and co-localized with high molecular mass DNA, probably the wasp chromosomes. Northern blot analysis of mRNAs extracted from the AA cell line showed transcription of some HdIV-encoded genes, although at low level. The effects of ecdysone treatment, HdIV re-infection and 42 °C heat-shock were analysed in the AA cell line. No effect was detected at the DNA (virus replication) or RNA (gene expression) levels, which may be due to the limitation of the present available tools.

scholarly journals Rearrangement and overexpression of the BCL-1/PRAD-1 gene in intermediate lymphocytic lymphomas and in t(11q13)-bearing leukemias

Blood ◽  
1993 ◽  
Vol 81 (11) ◽  
pp. 3063-3067 ◽  
Author(s):  
R Rimokh ◽  
F Berger ◽  
G Delsol ◽  
C Charrin ◽  
MF Bertheas ◽  
...  
Keyword(s):  
Northern Blot ◽  
Blot Analysis ◽  
Southern Blot Analysis ◽  
Northern Blot Analysis ◽  
Lymphoid Tissues ◽  
Over Expression ◽  
Lymphoid Neoplasia ◽  
Flanking Region ◽  
Variant Translocation

The t(11;14)(q13;q32) translocation and its molecular counterpart, BCL- 1 rearrangement, are consistent features of intermediate lymphocytic lymphoma (ILL). Rearrangement is thought to deregulate the nearby PRAD- 1/BCL-1 proto-oncogene that is a newly identified member of the cyclin family. To characterize further the association between rearrangement of chromosome 11q13 and over-expression of BCL-1. Southern blot analysis was performed in 33 cases of ILL, 5 cases of t(11;14)- associated leukemias, and 1 case of leukemia carrying a variant translocation t(11;19)(q13;q13) using three separate BCL-1 locus probes. When RNA was available, BCL-1 expression was assessed by Northern blot analysis. DNA from 19 of 33 ILL (57%) showed BCL-1 rearrangement, 16 involving the major translocation cluster (MTC) region and 3 involving a new breakpoint cluster located in the 5' flanking region of the BCL-1 gene. DNA from 3 of 6 t(11q13)-associated leukemias demonstrated a rearrangement involving the MTC. Northern blot analysis showed that BCL-1 was overexpressed in 14 of 15 ILL and in all leukemias analyzed (included the t(11;19) leukemia) relative to normal and malignant lymphoid tissues. These results constitute additional elements in favor of the role of BCL-1 in lymphoid neoplasia and allow us to speculate about its mechanisms of activation.

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