Cloning of Bna-miR1140 Gene Promoter in Rape and Preliminary Identification of Its Expression Pattern

doi:10.13560/j.cnki.biotech.bull.1985.2016.07.010

Abstract

Abstract: In order to explore the expression pattern and regulation mechanism of Bna-miR1140,the upstream 1.5 kb fragment from Bna-miR1140 precursor was cloned in a cultivated variety Westar of rape by PCR approach on the basis of rapeseed miRNAs chip results in our laboratory,and their cis-acting elements were analyzed. Then plant expression vector of GUS reporter genes was constructed,the vector miR1140 pro∷GUS was transformed into variety Westar by Agrobacterium-mediated approach,and 5 positive transgenic lines were obtained. GUS chemically staining the varied tissues of T1 generation groups of positive rape strains,the results showed that the upstream 1.5 kb region of miR1140 precursor sequence presented the function of promoter,driving GUS expression in rapeseed,moreover,only specific expression in the petiole and leaf axil,indicating that miR1140Pro was a specific promoter.

Key words: rapeseed, Bna-miR1140, promoter, expression pattern

DONG Yun, WANG Yi, JIN Feng-wei, SUN Wan-cang, LIU Zi-gang, FANG Yan, XU Miao-yun, WANG Lei. Cloning of Bna-miR1140 Gene Promoter in Rape and Preliminary Identification of Its Expression Pattern[J]. Biotechnology Bulletin, 2016, 32(7): 66-72.

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