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 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.

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 Insulin-like growth factor 1 (IGF-1) gene expression in porcine ovarian tissue

1993 ◽  
Vol 73 (2) ◽  
pp. 253-257 ◽  
Author(s):  
S. T. Charlton ◽  
B. J. Cameron ◽  
D. R. Glimm ◽  
G. R. Foxcroft ◽  
J. J. Kennelly
Keyword(s):  
Gene Expression ◽  
Growth Factor ◽  
Northern Blot ◽  
Blot Analysis ◽  
Ribonucleic Acid ◽  
Northern Blot Analysis ◽  
Ovarian Tissue ◽  
Messenger Ribonucleic Acid ◽  
Polyadenylated Rna ◽  
Mrna Transcripts

A study was conducted to demonstrate IGF-1 gene expression in porcine ovarian tissue. Using northern blot analysis, IGF-1 messenger ribonucleic acid (mRNA) transcripts could not be consistently detected in total RNA. Authentic mRNA transcripts were detected in polyadenylated RNA-enriched RNA with sizes of 8.0, 3.6 and 2.3 bases, and possibly 0.8–1.1 kbases. Key words: Ovary, IGF-1, pig, mRNA

Prolactin and growth hormone mRNA and protein characterization in SMtTW rat pituitary tumours

1995 ◽  
Vol 15 (3) ◽  
pp. 233-243 ◽  
Author(s):  
M Delhase ◽  
F Rajas ◽  
P Verdood ◽  
C Remy ◽  
P Chevallier ◽  
...  
Keyword(s):  
Western Blot ◽  
Northern Blot ◽  
Blot Analysis ◽  
Northern Blot Analysis ◽  
Biological Significance ◽  
Protein Characterization ◽  
Pituitary Tumours ◽  
Rat Pituitary ◽  
Normal Rat

ABSTRACT We have combined different techniques to analyse passages of five different rat spontaneous pituitary tumours (SMtTW) that were transplanted under the kidney capsule. These tumours were secreting prolactin (PRL), GH or both hormones. RIA, immunocytochemistry (ICC) and Western blot analysis were applied to characterize the hormone(s) stored (ICC and Western blot) and secreted (RIA). mRNA content was analysed by PCR, Northern blot analysis and in situ hybridization. The data point not only to the reliability of the techniques used at both protein and RNA levels for each tumour studied but also to the complementarity of some techniques. For example, whereas Northern blot analysis demonstrates the presence and size of hormone mRNA, in situ hybridization indicates the percentage of cells expressing a given hormone mRNA and allows the presence of one population (or more) of cells in a given tumour to be identified. Moreover, the tumours were compared with normal rat pituitary. Although the PRL and GH mRNAs were identical in size, the amount of mRNA was lower in the tumours. At the protein level, the PRL and GH variants exhibited a different pattern of expression in tumours compared with the normal rat pituitary. The biological significance of these differences is discussed.

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.

scholarly journals Expression and tissue distribution of rat sulfated glycoprotein-1 (prosaposin).

1996 ◽  
Vol 44 (4) ◽  
pp. 327-337 ◽  
Author(s):  
C R Morales ◽  
M El-Alfy ◽  
Q Zhao ◽  
S A Igdoura
Keyword(s):  
Epithelial Cells ◽  
Northern Blot ◽  
Blot Analysis ◽  
Northern Blot Analysis ◽  
Biological Fluids ◽  
Cell Types ◽  
Rat Tissues ◽  
Fluid Compartments ◽  
Rat Tissue ◽  
Sulfated Glycoprotein

Sulfated glycoprotein-1 (SGP-1/prosaposin) exists as a sulfated secreted protein or as a lysosomal precursor of four smaller saposin molecules. The protein exhibits ubiquitous expression, evolutionary conservation, and diverse tissue inducibility. The lysosomal form of SGP-1 plays a role in the hydrolysis of glycolipids and sphingomyelin. The function of the secreted form of SGP-1 is still unclear. However, it could act as a glycolipid transfer protein, since several gangliosides (a series) were found to bind with high affinity to prosaposin. To identify cell types that produce SGP-1 mRNA, we constructed an SGP-1 cDNA and used for screening of different rat tissues by Northern blot analysis. To localize the translation product of SGP-1 transcripts, we immunostained the same tissues with an anti-SGP-1 antibody. The SGP-1 cDNA construct was generated by amplifying a rat testicular Zap cDNA library by PCR (polymerase chain reaction) with two synthetic oligonucleotide primers. A positive signal of 1.7 KB was isolated, subcloned into the pGEM-7Zf (+). Sequence analysis showed a near-identical nucleotide and amino acid similarity to a previous rat SGP-1 cDNA. The majority of the heterogeneites were conservative substitutions. Northern blot analysis demonstrated that all examined rat tissue and organs have SGP-1 mRNA. Immunocytochemistry identified two staining patterns in the cytoplasm of positive cells: (a) a granular reaction characteristic of lysosomes in the supranuclear and basal regions of epithelial cells and in the perinuclear region of neurons; and (b) a homogeneous reaction in the cytoplasm of Sertoli cells, Type II pneumocytes, macrophages, and epithelial cells lining the choroid plexus. The latter staining pattern could be characteristic of cells that exhibit a secretory routing of SGP- 1. The production of SGP-1 by a variety of specialized cells lining fluid compartments suggests that its secreted form has a role in the transport of lipids in biological fluids, possibly by the formation of soluble complexes with glycolipids. Similarly, the lysosomal form of SGP-1/prosaposin and their derived saposins also solubilizes certain glycolipids to promote their degradation by specific hydrolases.

scholarly journals Tissue- and cell-specific expression of Ins(1,4,5)P3 3-kinase isoenzymes

1995 ◽  
Vol 306 (2) ◽  
pp. 429-435 ◽  
Author(s):  
V Vanweyenberg ◽  
D Communi ◽  
C S D'Santos ◽  
C Erneux
Keyword(s):  
Cell Lines ◽  
Northern Blot ◽  
Blot Analysis ◽  
Northern Blot Analysis ◽  
Biological Data ◽  
Human Neuroblastoma Cell ◽  
Rat Tissues ◽  
Human Neuroblastoma ◽  
Mrna Species ◽  
Kinase A

The phosphorylation of Ins(1,4,5)P3 (InsP3) to Ins(1,3,4,5)P4 (InsP4) is catalysed by InsP3 3-kinase. Molecular-biological data have shown the presence of two human isoenzymes of InsP3 3-kinase, namely InsP3 3-kinases A and B. We have isolated from a rat thymus cDNA library a 2235 bp cDNA (clone B15) encoding rat InsP3 3-kinase B. Northern-blot analysis of mRNA isolated from rat tissues (thymus, testis, brain, spleen, liver, kidney, heart, lung and intestine) revealed that a rat InsP3 3-kinase B probe hybridized to a 6 kb mRNA in lung, thymus, testis, brain and heart. In contrast, Northern-blot analysis of the same tissues probed under stringent conditions with a rat InsP3 3-kinase A probe hybridized to a 2 kb mRNA only in brain and a 1.8-2.0 kb mRNA species in testis. Northern-blot analysis of three human cell lines (HL-60, SH-SY5Y and HTB-138) probed with a human InsP3 3-kinase B probe showed the presence of a 6 kb mRNA in all cell lines, except in the human neuroblastoma cell line (SH-SY5Y), where two mRNA species of 5.7 and 6 kb were detected. Using the same blot, no hybridization signal could be seen with a human InsP3 3-kinase A probe. Altogether, our data are consistent with the notion that the two InsP3 3-kinase isoenzymes, A and B, are specifically expressed in different tissues and cells.

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.

Sign in / Sign up

Export Citation Format

Share Document