Differential screening of a PCR-generated mouse embryo cDNA library: glucose transporters are differentially expressed in early postimplantation mouse embryos

Development ◽  
1992 ◽  
Vol 116 (3) ◽  
pp. 555-561 ◽  
Author(s):  
D.E. Smith ◽  
T. Gridley
Keyword(s):  
Cdna Library ◽  
Pcr Amplification ◽  
Glucose Transporters ◽  
Distal Portion ◽  
Mouse Embryos ◽  
Differential Screening ◽  
Differentially Expressed ◽  
Nucleotide Sequence Analysis ◽  
Cdna Clones ◽  
Glut 1

Differential screening of a cDNA library constructed using PCR amplification techniques from RNA isolated from the distal portion (embryonic ectoderm, mesoderm and visceral endoderm) of 7.5 days post coitum (dpc) mouse embryos led to the isolation of two cDNA clones expressed at higher levels in 7.5 dpc embryos than 12.5 dpc embryos. Nucleotide sequence analysis revealed that each of these clones was a different member of the family of facilitative glucose transporters (Glut genes). The differentially expressed cDNA clones represent mouse Glut-1 and Glut-3. Levels of the Glut-3 mRNA declined 14-fold between days 7.5 and 12.5 of gestation, and were under our limits of detection by 14.5 dpc. The levels of the Glut-1 mRNA declined about 3-fold between days 7.5 and 12.5 of gestation. Analysis of the expression of these genes by in situ hybridization revealed striking differences in transcript localization in early postimplantation mouse embryos. At 7.5 dpc, both transporters were expressed more strongly in extraembryonic tissues than in the embryo proper. While both transporters were expressed in the amnion and chorion, only Glut-1 was expressed in the ectoplacental cone. In the yolk sac, Glut-3 appeared to be expressed only in the endoderm while Glut-1, although expressed in both layers, was expressed more strongly in the mesoderm layer. Thus, the two transporters have relatively reciprocal sites of expression in the developing extraembryonic membranes. Expression of Glut-1 was fairly widespread in the embryo at 8.5 dpc, but by 10.5 dpc expression was down-regulated and was observed in the eye and the spinal cord.(ABSTRACT TRUNCATED AT 250 WORDS)

Construction of an equalized cDNA library fromColletotrichum lagenariumand its application to the isolation of differentially expressed genes

2000 ◽  
Vol 46 (2) ◽  
pp. 150-158 ◽  
Author(s):  
Atsuko Inagaki ◽  
Yoshitaka Takano ◽  
Yasuyuki Kubo ◽  
Kazuyuki Mise ◽  
Iwao Furusawa
Keyword(s):  
Amino Acid ◽  
Cdna Library ◽  
Differentially Expressed Genes ◽  
Kinetic Method ◽  
Differential Screening ◽  
Differentially Expressed ◽  
Phytopathogenic Fungus ◽  
Cdna Clones ◽  
Appressorium Formation ◽  
Colletotrichum Lagenarium

To establish an efficient screening system for differentially expressed genes of a phytopathogenic fungus Colletotrichum lagenarium, we constructed an equalized (normalized) cDNA library from C. lagenarium and used this library for differential screening. For the isolation of genes involved in infection-related developments of conidia, conidia undergoing appressorium differentiation were selected as the source of materials for construction of the cDNA library. The equalization of cDNA was performed twice using a kinetic method, and the products were cloned into a plasmid vector. Colony hybridization with nine probes of different abundance showed a reduction in abundance variation from at least 276-fold in the original library to 10-fold in the equalized cDNA library, which demonstrated that the cDNA was successfully equalized. By differential hybridization of 1900 cDNA clones in the equalized cDNA library and RNA blot analysis of candidate clones, we identified 11 independent cDNA clones, designated CAD1 through CAD11, that were expressed in appressorium-differentiating conidia, but not in vegetative mycelia. The transcripts of CAD1 and CAD2 hardly accumulated in preincubated conidia, whereas those of CAD3 and CAD4 accumulated highly and slightly, respectively. The amount of the four CAD transcripts increased at the early stage of the appressorium formation process. Sequence analysis of CAD1 revealed that CAD1 would encode for 101 amino acid polypeptides, which showed homology to metallothioneins. Deduced amino acid sequence of CAD2 would encode 278 amino acid polypeptides, and showed high homology to genes in aflatoxin, and sterigmatocystin gene clusters of Aspergillus parasiticus and A. nidulans, respectively. Key words: equalized cDNA library, differential screening, Colletotrichum lagenarium, appressorium formation, CAD genes.

Low-Flow Ischemia Leads to Translocation of Canine Heart GLUT-4 and GLUT-1 Glucose Transporters to the Sarcolemma In Vivo

Circulation ◽  
1997 ◽  
Vol 95 (2) ◽  
pp. 415-422 ◽  
Author(s):  
Lawrence H. Young ◽  
Yin Renfu ◽  
Raymond Russell ◽  
Xiaoyue Hu ◽  
Michael Caplan ◽  
...  
Keyword(s):  
Glucose Transporters ◽  
Low Flow ◽  
Canine Heart ◽  
Glut 1 ◽  
Glut 4

scholarly journals Differential Screening of a Subtracted cDNA Library: A Method to Search for Genes Preferentially Expressed in Multiple Tissues

BioTechniques ◽  
1997 ◽  
Vol 23 (6) ◽  
pp. 1084-1086 ◽  
Author(s):  
H. Jin ◽  
X. Cheng ◽  
L. Diatchenko ◽  
P.D. Siebert ◽  
C.-C. Huang
Keyword(s):  
Cdna Library ◽  
Differential Screening

scholarly journals Expression of Glucose Transporters 1 and 3 in Metastatic and Non-Metastatic Lower Lip Squamous Cell Carcinoma

2014 ◽  
Vol 25 (5) ◽  
pp. 372-378 ◽  
Author(s):  
Clarissa Favero Demeda ◽  
Cyntia Helena Pereira de Carvalho ◽  
Ana Rafaela Luz de Aquino ◽  
Cassiano Francisco Weege Nonaka ◽  
Lélia Batista de Souza ◽  
...  
Keyword(s):  
Squamous Cell Carcinoma ◽  
Cell Carcinoma ◽  
Glucose Uptake ◽  
Squamous Cell ◽  
Histological Grade ◽  
Glucose Transporters ◽  
Nodal Metastasis ◽  
Invasive Front ◽  
Lower Lip ◽  
Glut 1

This study aimed to evaluate the immunoexpression of glucose transporters 1 (GLUT-1) and 3 (GLUT-3) in metastatic and non-metastatic lower lip squamous cell carcinoma (LLSCC). Twenty LLSCCs with regional nodal metastasis and 20 LLSCCs without metastasis were selected. The distribution of staining and the percentage of GLUT-1 and GLUT-3 staining in each tumor core and at the deep invasive front were assessed. Most tumors (70%) exhibited peripheral staining for GLUT-1 in nests, sheets and islands of neoplastic cells, whereas predominantly central staining was observed for GLUT-3 (72.5%). A high percentage of GLUT-1-positive cells was observed at the deep invasive front and in the tumor core of metastatic and non-metastatic tumors (p>0.05). The percentage of GLUT-1-positive cells was much higher than that of GLUT-3-positive cells both in the deep invasive front (p<0.001) and in the tumor core (p<0.001) of LLSCCs. No significant differences in the percentage of GLUT-1- and GLUT-3-positive cells were observed according to nodal metastasis, clinical stage or histological grade of malignancy (p>0.05). In conclusion, the results of the present study suggest an important role of GLUT-1 in glucose uptake in LLSCCs, although this protein does not seem to be involved in the progression of these tumors. On the other hand, GLUT-3 expression may represent a secondary glucose uptake mechanism in LLSCCs.

Construction and Identification by Partial Nucleotide Sequence Analysis of Bovine Casein and β-Lactoglobulin cDNA Clones

DNA ◽  
1982 ◽  
Vol 1 (4) ◽  
pp. 375-386 ◽  
Author(s):  
I.M. WILLIS ◽  
A.F. STEWART ◽  
A. CAPUTO ◽  
A.R. THOMPSON ◽  
A.G. MACKINLAY
Keyword(s):  
Sequence Analysis ◽  
Nucleotide Sequence ◽  
Nucleotide Sequence Analysis ◽  
Cdna Clones ◽  
Partial Nucleotide Sequence ◽  
Β Lactoglobulin

scholarly journals Isolation of cDNA clones for genes exhibiting reduced expression after differentiation of murine teratocarcinoma stem cells.

1984 ◽  
Vol 4 (10) ◽  
pp. 2142-2150 ◽  
Author(s):  
R A Levine ◽  
G J LaRosa ◽  
L J Gudas
Keyword(s):  
Stem Cells ◽  
Retinoic Acid ◽  
Cyclic Amp ◽  
Cdna Library ◽  
In Vitro Transcription ◽  
In Vitro Translation ◽  
Transcriptional Level ◽  
Cdna Clones ◽  
Dibutyryl Cyclic Amp

In the absence of retinoic acid, PSA-G teratocarcinoma stem cells spontaneously differentiate at a moderate frequency into fibroblast-like cells. In the presence of retinoic acid and dibutyryl cyclic AMP, PSA-G stem cells differentiate into parietal endoderm cells. We prepared a cDNA library from undifferentiated PSA-G teratocarcinoma stem cells; this cDNA library was then screened for gene sequences which exhibit a reduction in expression during the differentiation of these stem cells. From ca. 1,000 clones screened, eight independent sequences were isolated. The level of expression of these cloned genes decreases by 3.0-fold to more than 10-fold after differentiation of PSA-G cells into fibroblast-like cells. After treatment of either PSA-G or F9 teratocarcinoma cells with retinoic acid and dibutyryl cyclic AMP for 72 h, the expression of seven genes is inhibited by two- to fourfold. This decrease of clone-specific transcripts can be detected within 12 h after the addition of retinoic acid. Hybridization-selection and in vitro translation experiments identified the proteins encoded by three of the cloned genes: pST 6-23 codes for a 89,000-dalton protein, pST 7-105 codes for a 41,000-dalton protein, and pST 9-31 codes for a 34,000-dalton protein. The 89,000-dalton protein encoded by pST 6-23 is a heat shock protein. In vitro transcription experiments demonstrate that the retinoic acid-mediated decrease in pST 6-135- and pST 1-68-specific RNA occurs at the transcriptional level and that dibutyryl cyclic AMP acts posttranscriptionally to further depress the levels of these RNAs.

Isolation of Wound-Specific cDNA Clones from a cDNA Library Prepared with mRNAs of Alkali-Burned Rabbit Corneas

Cornea ◽  
1991 ◽  
Vol 10 (4) ◽  
pp. 322-329 ◽  
Author(s):  
Takeshi Haseba ◽  
Mitsuru Nakazawa ◽  
Winston W-Y. Kao ◽  
Ramesh Murthy ◽  
Candace W-C. Kao
Keyword(s):  
Cdna Library ◽  
Cdna Clones
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