Construction and analysis of SSH library from ultra-dwarf mutant in upland cotton

The total cotton RNA of upland cotton ultra-dwarf mutant(tester)and its wild-type near-isogenic line M104(driver) was isolated using SSH(suppression subtractive hybridization) to establish differential expression cDNA library.The reverse Northern blot was conducted with cDNA of ultra-dwarf mutant or M104 alone as probes,to screen differential expression clones and analyze the sequence similarity.2 530 clones were obtained from the cDNA library.Totally 668 differential expression clones out of 2 530 were screened through the reverse Northern blot.After sequencing,539 EST sequences were got,including 122 unigenes,36 contigs and 86 singletons,with the average length of 1 319.26 bp.These genes were related to regulation of metabolism and secondary metabolism,transcriptional regulation,biosynthesis,cell apoptosis and signal transduction.So,the construction and analysis of SSH library might provide a foundation for further research on structure and function of gene related to ultra-dwarf mutation in upland cotton.