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 Construction and Differential Screening of a cDNA Library Specific to Osmotic Stress of Haloxylon ammodendron Seedlings

BMB Reports ◽  
2004 ◽  
Vol 37 (5) ◽  
pp. 527-532 ◽  
Author(s):  
Xiao-Cheng Jiang ◽  
Xin-Hong Guo ◽  
Xiao-Ling Pan ◽  
Song-Quan Song
Keyword(s):  
Osmotic Stress ◽  
Cdna Library ◽  
Differential Screening ◽  
Haloxylon Ammodendron

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.

Isolation of Novel and Known Genes from a Human Fetal Cochlear cDNA Library Using Subtractive Hybridization and Differential Screening

Genomics ◽  
1994 ◽  
Vol 23 (1) ◽  
pp. 42-50 ◽  
Author(s):  
Nahid G. Robertson ◽  
Umang Khetarpal ◽  
Gustavo A. Gutiérrez-Espeleta ◽  
Frederick R. Bieber ◽  
Cynthia C. Morton
Keyword(s):  
Cdna Library ◽  
Subtractive Hybridization ◽  
Differential Screening

scholarly journals 347 ETHYLENE-INDUCED GENE EXPRESSION DURING FLOWER PETAL ABSCISSION OF ZONAL GERANIUM

HortScience ◽  
1994 ◽  
Vol 29 (5) ◽  
pp. 480d-480
Author(s):  
David G. Clark ◽  
Kathleen B. Evensen
Keyword(s):  
Gene Expression ◽  
Cdna Library ◽  
Plant Development ◽  
Differential Screening ◽  
Flower Petal ◽  
Pelargonium Xhortorum ◽  
Ethylene Action ◽  
Regulated Gene Expression ◽  
Identification And Characterization

Ethylene-regulated gene expression is being studied in several plant systems, but the exact mechanism of ethylene action during plant development and senescence is poorly understood. When geranium (Pelargonium Xhortorum) flowers are exposed to 1 μ1/L of ethylene gas for 1 hour, petals begin to abscise within 60-90 minutes from the start of treatment, The rapidity of the response implies that it must be very direct. We now demonstrate that ethylene acts at the level of message accumulation. We have constructed a cDNA library from mRNA isolated from ethylene-treated geranium gynoecia. Ethylene-induced clones have been isolated by differential screening of this library with cDNA probes synthesized from ethylene-treated and untreated geranium gynoecia mRNA. Identification and characterization of these clones will be discussed.

scholarly journals Cloning by differential screening of a Xenopus cDNA that encodes a kinesin-related protein.

1991 ◽  
Vol 11 (6) ◽  
pp. 3395-3398 ◽  
Author(s):  
R Le Guellec ◽  
J Paris ◽  
A Couturier ◽  
C Roghi ◽  
M Philippe
Keyword(s):  
Aspergillus Nidulans ◽  
Cdna Library ◽  
Nuclear Division ◽  
Differential Screening ◽  
Related Protein ◽  
Xenopus Egg ◽  
Characteristic Features

By differential screening of a Xenopus egg cDNA library, we selected nine clones (Eg1 to Eg9) corresponding to mRNAs which are deadenylated and released from polysomes soon after fertilization. The sequence of one of these clones (Eg5) revealed that the corresponding protein has the characteristic features of a kinesin-related protein. More specifically, Eg5 was found to be nearly 30% identical to a kinesin-related protein encoded by bimc, a gene involved in nuclear division in Aspergillus nidulans.

Differential screening of a cDNA library with cDNA probes amplified in a heterologous host: isolation of murine GRP78 (BiP) and other serum-regulated low-abundance mRNAs

Gene ◽  
1989 ◽  
Vol 82 (2) ◽  
pp. 291-303 ◽  
Author(s):  
Craig L.J. Parfett ◽  
Reinhold Hofbauer ◽  
Katarzyna Brudzynski ◽  
Dylan R. Edwards ◽  
David T. Denhardt
Keyword(s):  
Cdna Library ◽  
Differential Screening ◽  
Heterologous Host

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)

Cloning by differential screening of a Xenopus cDNA that encodes a kinesin-related protein

1991 ◽  
Vol 11 (6) ◽  
pp. 3395-3398
Author(s):  
R Le Guellec ◽  
J Paris ◽  
A Couturier ◽  
C Roghi ◽  
M Philippe
Keyword(s):  
Aspergillus Nidulans ◽  
Cdna Library ◽  
Nuclear Division ◽  
Differential Screening ◽  
Related Protein ◽  
Xenopus Egg ◽  
Characteristic Features

By differential screening of a Xenopus egg cDNA library, we selected nine clones (Eg1 to Eg9) corresponding to mRNAs which are deadenylated and released from polysomes soon after fertilization. The sequence of one of these clones (Eg5) revealed that the corresponding protein has the characteristic features of a kinesin-related protein. More specifically, Eg5 was found to be nearly 30% identical to a kinesin-related protein encoded by bimc, a gene involved in nuclear division in Aspergillus nidulans.

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