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A preliminary study on the genetic transformation system of plant gene overexpression based on a mCherry fluorescent protein vector

  

  1. (1 College of Chemistry and Life Sciences, Zhejiang Normal University, Jinhua 321004, China; 2 Institute of Virology and Biotechnology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China)
  • Online:2015-11-25 Published:2015-12-05

Abstract:

Abstract: Overexpression is an important approach for functional characterization of plant genes and genetic improvement of crop plants. To improve the efficiency of characterization of transgenic plants, a transformation vector with the maize (Zea mays) ubiquitin\|1 promoter (P\|Ubi) and the pea (Pisum sativum) T3A\|polyA sequence in the overexpression cassette was constructed using the mCherry red fluorescent protein as a reporter. The coding sequences of four receptor\|like kinases of rice (Oryza sativa) were inserted into the overexpression cassette of the vector and then transformed into the rice cultivar Taijing\|394, respectively. mCherry fluorescence assay of the T1 transgenic seeds showed that on average, 625% of the T1 lines segregated with a 3∶1 ratio. Based on the results of real\|time quantitative reverse\|transcription polymerase chain reaction assays, the relative gene\|expression levels of the overexpressed genes in the transgene\|positive plants were significantly higher than those of the genes in the negative segregant and untransformed recipient cultivar. The vector system can be used for large\|scale screening of transgenic progeny and obtaining of transgenic lines with target genes overexpressed, so as to promote the functional characterization of plant genes and the application of transgenic technology.

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