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