ÃÞ »¨ ѧ ±¨ Cotton Science 2009£¬21(3)£º190-195
Cloning and Sequence Analysis of Fiber-specific Expressional Promoter GhCesA4 from Cotton( Gossypium hirsutum ) Abstract£ºDifferentiation of cotton fiber experience four development stages, including initiation, prolongation, secondary wall thickening, and dehydration and maturation. During the stages, many genes, known to be fiber-specific or preferentially expressed in cotton fiber, are developmentally expressed. Inhere promoter of cotton fiber development related gene play an important role. In this experiment, we cloned cotton fiber-specific promoter GhCes A4 from "Xinluzao 19", GenBank accession number is EU183119. The promoter is cloned into pMD 19-T vector, then we successfully identified and obtained recombinant plasmid pMD 19-T/ Ghces A4 through vector specific PCR primer M13-47 and RV-M. Sequencing and sequence analysis showed the promoter is up of 1503 bp nucleotide and has 98% identity with GhCes A4 gene submitted in GenBank. Respectively restriction enzyme Cla ¢ñ and Bam H ¢ñ digested recombinant plasmid pMD 19-T/ GhCes A4 and binary plant expression vector pBI121,recovering the small fragment in the recombinant plasmid pMD 19-T/ GhCes A4 and the big fragment lack of CaMV35S promoter in binary plant expression vector pBI121.By ligating, transforming, digestion and sequencing, we obtained a new pattern plant expression vector which was driven by GhCes A4 GUS reporter gene, namely pBI- GhCes A4.
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