Identification of differentially expressed genes in developing cotton fibers (Gossypium hirsutum L) through differential display

Cotton fibers are differentiated, non-dividing cells that originate from the epidermal layer of developing ovules. To identify genes involved in cotton fiber development, we performed non-radioactive differential display reverse transcriptase PCR (DDRT-PCR) on the purified mRNA. This technique was t...

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Autores principales:	Iqbal,Saima, Bashir,Aftab, Naseer,Hafiza Masooma, Ahmed,Moddassir, Malik,Kauser A
Lenguaje:	English
Publicado:	Pontificia Universidad Católica de Valparaíso 2008
Materias:	cotton fiber differentially expressed transcripts Gossypium hirsutum non-radioactive
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582008000100005
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spelling	oai:scielo:S0717-345820080001000052008-10-28Identification of differentially expressed genes in developing cotton fibers (Gossypium hirsutum L) through differential displayIqbal,SaimaBashir,AftabNaseer,Hafiza MasoomaAhmed,ModdassirMalik,Kauser A cotton fiber differentially expressed transcripts Gossypium hirsutum non-radioactive Cotton fibers are differentiated, non-dividing cells that originate from the epidermal layer of developing ovules. To identify genes involved in cotton fiber development, we performed non-radioactive differential display reverse transcriptase PCR (DDRT-PCR) on the purified mRNA. This technique was tested on mRNA isolated from five different developmental stages of cotton fiber including 0, 5, 10, 15 and 20 DPA (days after pollination). The mRNA purified from total RNA was reversibly transcribed using three anchored oligo-dT primers. Polymerase chain reaction (PCR) amplification of each cDNA preparation was carried out in combination with seven arbitrary primers. The amplified products were resolved on 1% agarose gel containing ethidium bromide. DNA was extracted from seventeen differentially expressed bands and cloned in pTZ57R/T vector. The sequencing and BLAST search analysis indicated that 12 of the differentially expressed genes matched the previously characterized genes, while 3 of them matched the uncharacterized sequences of cotton fiber expressed sequence tags (ESTs) reported previously to be associated with cotton fiber and 2 of the clones had homology with putative proteins. The technique can be used to efficiently identify differentially expressed genes and can be expanded to large scale studies by increasing the number of random decamers.info:eu-repo/semantics/openAccessPontificia Universidad Católica de ValparaísoElectronic Journal of Biotechnology v.11 n.1 20082008-01-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582008000100005en10.4067/S0717-34582008000100005
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	cotton fiber differentially expressed transcripts Gossypium hirsutum non-radioactive
spellingShingle	cotton fiber differentially expressed transcripts Gossypium hirsutum non-radioactive Iqbal,Saima Bashir,Aftab Naseer,Hafiza Masooma Ahmed,Moddassir Malik,Kauser A Identification of differentially expressed genes in developing cotton fibers (Gossypium hirsutum L) through differential display
description	Cotton fibers are differentiated, non-dividing cells that originate from the epidermal layer of developing ovules. To identify genes involved in cotton fiber development, we performed non-radioactive differential display reverse transcriptase PCR (DDRT-PCR) on the purified mRNA. This technique was tested on mRNA isolated from five different developmental stages of cotton fiber including 0, 5, 10, 15 and 20 DPA (days after pollination). The mRNA purified from total RNA was reversibly transcribed using three anchored oligo-dT primers. Polymerase chain reaction (PCR) amplification of each cDNA preparation was carried out in combination with seven arbitrary primers. The amplified products were resolved on 1% agarose gel containing ethidium bromide. DNA was extracted from seventeen differentially expressed bands and cloned in pTZ57R/T vector. The sequencing and BLAST search analysis indicated that 12 of the differentially expressed genes matched the previously characterized genes, while 3 of them matched the uncharacterized sequences of cotton fiber expressed sequence tags (ESTs) reported previously to be associated with cotton fiber and 2 of the clones had homology with putative proteins. The technique can be used to efficiently identify differentially expressed genes and can be expanded to large scale studies by increasing the number of random decamers.
author	Iqbal,Saima Bashir,Aftab Naseer,Hafiza Masooma Ahmed,Moddassir Malik,Kauser A
author_facet	Iqbal,Saima Bashir,Aftab Naseer,Hafiza Masooma Ahmed,Moddassir Malik,Kauser A
author_sort	Iqbal,Saima
title	Identification of differentially expressed genes in developing cotton fibers (Gossypium hirsutum L) through differential display
title_short	Identification of differentially expressed genes in developing cotton fibers (Gossypium hirsutum L) through differential display
title_full	Identification of differentially expressed genes in developing cotton fibers (Gossypium hirsutum L) through differential display
title_fullStr	Identification of differentially expressed genes in developing cotton fibers (Gossypium hirsutum L) through differential display
title_full_unstemmed	Identification of differentially expressed genes in developing cotton fibers (Gossypium hirsutum L) through differential display
title_sort	identification of differentially expressed genes in developing cotton fibers (gossypium hirsutum l) through differential display
publisher	Pontificia Universidad Católica de Valparaíso
publishDate	2008
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582008000100005
work_keys_str_mv	AT iqbalsaima identificationofdifferentiallyexpressedgenesindevelopingcottonfibersgossypiumhirsutumlthroughdifferentialdisplay AT bashiraftab identificationofdifferentiallyexpressedgenesindevelopingcottonfibersgossypiumhirsutumlthroughdifferentialdisplay AT naseerhafizamasooma identificationofdifferentiallyexpressedgenesindevelopingcottonfibersgossypiumhirsutumlthroughdifferentialdisplay AT ahmedmoddassir identificationofdifferentiallyexpressedgenesindevelopingcottonfibersgossypiumhirsutumlthroughdifferentialdisplay AT malikkausera identificationofdifferentiallyexpressedgenesindevelopingcottonfibersgossypiumhirsutumlthroughdifferentialdisplay
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Identification of differentially expressed genes in developing cotton fibers (Gossypium hirsutum L) through differential display

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