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Identification of differentially expressed genes in developing cotton fibers (Gossypium hirsutum L) through differential display
Iqbal, Saima; Bashir, Aftab; Naseer, Hafiza Masooma; Ahmed, Moddassir; Malik, Kauser A.
Afiliação
  • Iqbal, Saima; National Institute for Biotechnology and Genetic Engineering. Plant Biotechnology Division. Faisalabad. PK
  • Bashir, Aftab; National Institute for Biotechnology and Genetic Engineering. Plant Biotechnology Division. Faisalabad. PK
  • Naseer, Hafiza Masooma; National Institute for Biotechnology and Genetic Engineering. Plant Biotechnology Division. Faisalabad. PK
  • Ahmed, Moddassir; National Institute for Biotechnology and Genetic Engineering. Plant Biotechnology Division. Faisalabad. PK
  • Malik, Kauser A; Food and Agriculture Planning Commission. Islamabad. PK
Electron. j. biotechnol ; 11(1): 50-59, Jan. 2008. ilus, tab
Artigo em Inglês | LILACS | ID: lil-522160
Biblioteca responsável: CL1.1
ABSTRACT
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 percent 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.
Assuntos
Texto completo: Disponível Coleções: Bases de dados internacionais Base de dados: LILACS Assunto principal: Óvulo / Diferenciação Celular / Gossypium herbaceum / Fibra de Algodão Tipo de estudo: Estudo diagnóstico Idioma: Inglês Revista: Electron. j. biotechnol Assunto da revista: Biotecnologia Ano de publicação: 2008 Tipo de documento: Artigo País de afiliação: Paquistão Instituição/País de afiliação: Food and Agriculture Planning Commission/PK / National Institute for Biotechnology and Genetic Engineering/PK
Texto completo: Disponível Coleções: Bases de dados internacionais Base de dados: LILACS Assunto principal: Óvulo / Diferenciação Celular / Gossypium herbaceum / Fibra de Algodão Tipo de estudo: Estudo diagnóstico Idioma: Inglês Revista: Electron. j. biotechnol Assunto da revista: Biotecnologia Ano de publicação: 2008 Tipo de documento: Artigo País de afiliação: Paquistão Instituição/País de afiliação: Food and Agriculture Planning Commission/PK / National Institute for Biotechnology and Genetic Engineering/PK
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