Data/hora: 02/06/2024 04:54:51

Provedor de dados: | Electronic Journal of Biotechnology
País: | Chile
Título: | Identification of differentially expressed genes in developing cotton fibers (Gossypium hirsutum L) through differential display
Autores: | Iqbal,Saima; Bashir,Aftab; Naseer,Hafiza Masooma; Ahmed,Moddassir; Malik,Kauser A.
Data: | 2008-01-01
Ano: | 2008
Palavras-chave: | Cotton fiber; Differentially expressed transcripts; Gossypium hirsutum; Non-radioactive.
Resumo: | 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.
Tipo: | Journal article
Idioma: | Inglês
Identificador: | http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582008000100005
Editor: | Pontificia Universidad Católica de Valparaíso
Formato: | text/html
Fonte: | Electronic Journal of Biotechnology  v.11 n.1 2008