Original£ºCotton Science£¬2006£¬18(2)£º67-73

 

 

Cloning and Characterization of a Novel Cotton Fiber
Expressed Protein ( GhCFE ) cDNA

GUO Ying, GUO Wang-zhen*£¬ZHANG Tian-zhen
(State Key Laboratory of Crop Genetics & Germplasm Enhancement, Nanjing Agricultural University, Nanjing 210095, China )

Abstract£ºThrough randomly sequencing the cotton fiber cDNA library from 7235 germplasm line with elite fiber quality in Gossypium hirsutum L., a cDNA clone encoding cotton fiber expressed protein was isolated, designated as GhCFE ] (GenBank accession number: DQ073045). The full length of this cDNA clone was 1274 bp, with the open reading frame encoded 332 amino acids. The putative protein of this gene had an isoelectric point of 6.14 and a calculated molecular weight of 37.7 kD. It had a signal peptide with 27 amino acid residues in N-terminal. RT-PCR analysis indicated this gene did not express in root or stem, only had a weak expression in leaf, while it could be detected in ovules and fiber cells of different developmental stages, especially in elongating fibers. Phylogenetic tree showed GhCFE had high homologies with other CFEs reported previously. We cloned GhCFE into the fission yeast ( S. pombe ) vector pREP5N and detected that this gene had no significant effect on elongating cells or thickening cell wall in the transformed yeast. Further, sense expression vector containing 35S promoter and both sense and antisense expression vectors containing E6 promoter using pBI121 plasmid were constructed, and the work of transferring these recombined vectors into Gossypium hirsutum L. by Agrobacterium tumefaciens is ongoing.
Key words£ºGossypium hirsutum L.; fiber expressed protein; expression vector    [Full Text,3062KB]