Differentiation 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 GhCesA4 from "Xinluzao 19", GenBank accession number is EU183119. The promoter is cloned into pMD19-T vector, then we successfully identified and obtained recombinant plasmid pMD19-T/GhcesA4 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 GhCesA4 gene submitted in GenBank. Respectively restriction enzyme ClaⅠand BamHⅠdigested recombinant plasmid pMD19-T/GhCesA4 and binary plant expression vector pBI121,recovering the small fragment in the recombinant plasmid pMD19-T/GhCesA4 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 GhCesA4 GUS reporter gene, namely pBI-GhCesA4.
Key words
cotton
/
fiber-specific promoter /
GhCesA4 /
cloning /
sequence analysis
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