Establishment and Identification of a Normalized Full-Length cDNA Library of CCRI 36

Abstract

Short season cotton (SSC) breeding plays an important role in solving the competition for more growing area between cotton and food crops. To explore the premature mechanism of SSC, to clone genes that have a close relation with premature flowering, and to speed process of premature cultivar breeding, we established a normalized full-length cDNA library using the flower and bud of CCRI 36 by DSN (duplex-specific nuclease)-normalization method combined with SMART (switching mechanism at 5' end of RNA transcript) technique. The titer of un-amplified cDNA library was about 1.7×10 6 cfu mL −1 . The average size of cDNA inserts was 1 200 bp with a recombination rate of 100%. The abundance of transcripts Histon3 and UBQ7 decreased obviously in normalized cDNA library comparing with that in non-normalized samples detected by Virtual Northern Blot. Meanwhile, protein genes associated with flower development were obtained on the basis of the positive signal of cDNA library by PCR. These results indicated that the normalized full-length cDNA library has been established successfully, which is convenient for further study on the molecular mechanism and gene cloning of flower development.