Ubi1 intron sequence capable of enhancing gene expression and applications

A technology of gene expression and gene expression level, applied in the field of plant genetic engineering, can solve problems that have not been reported yet

Inactive Publication Date: 2012-10-24
THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI
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  • Abstract
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Problems solved by technology

Wang Yuebing compared the first intron of maize ubiquitin protein gene (ubi1), the first intron of rice actin gene (act1), the first intron of maize alcohol dehydrogenase gene (adh1) and potato The second intron (SBgLR2) of the high lysine gene SBgLR gene has the enhanced effect on gene expression, and it was found that ubi1 has the strongest ability to enhance the expression of the reporter gene; although it is known that not all sequences in the intron are effective in improving gene expression Necessary, but there is no report on the study of ubi1 intron modification by deletion analysis

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  • Ubi1 intron sequence capable of enhancing gene expression and applications
  • Ubi1 intron sequence capable of enhancing gene expression and applications
  • Ubi1 intron sequence capable of enhancing gene expression and applications

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Embodiment

[0070] 1. Construction of deletion vector pSG(13i-Pn)N

[0071] The intron deletion fragment was obtained by PCR amplification method, the amplification primers are shown in Table 4, and the PCR amplification diagram is shown in figure 1 . Pst I restriction sites are added to both ends of the amplified fragment. Since the 23-28bp of the CaMV35S promoter contains a Pst I restriction site, the intron sequence cannot be directly replaced on the pSG(+13i)N vector. During construction, the pSG(+13i)N vector (preserved by the Crop Genomics and Genetic Improvement Laboratory, Institute of Biotechnology, Chinese Academy of Agricultural Sciences) was first digested with Bam HI and Sac I, and the GUS+ubi1 fragment was recovered, and then inserted with the same double digestion The intermediate vector p7ZGUS(+13i) was constructed on the multiple cloning site of the obtained pGEM7Z vector (commercialized vector, which can be purchased in biological companies), and then the sequenced cor...

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Abstract

The invention discloses a ubi1 intron sequence capable of enhancing gene expression and applications. According to the sequence and applications, deletion analysis is conducted on a ubiquitin first intron from maize, the novel intron sequence capable of enhancing the foreign gene expression is obtained, the length of the intron sequence is 872bp, the adenine-thymine (AT) content is 61.0%, the gene expression can be improved by 13 times, and compared with the full-length ubiquitin first intron, the intron sequence improves the foreign gene expression ability by 30%. The intron obtained through the sequence can be used for constructing an efficient plant expression vector and improving the foreign gene expression level of a transgenic plant. According to the sequence and applications, the plant expression vector containing the modified intron is constructed, and the intron is used for cultivating of the transgenic plant to improve the expression of other genes in the transgenic plant.

Description

Technical field: [0001] The invention belongs to the technical field of plant genetic engineering and relates to a ubi1 intron sequence for enhancing gene expression. The present invention also relates to the application of the ubil intron. Background technique: [0002] Most eukaryotic genes are interrupted by one or more introns, these spacers can be transcribed into pre-mRNA, and these transcripts can only form mature after the intron fragments are removed by the splicing complex in the nucleus mRNA, which then travels through the nuclear pore into the cytoplasm for translation. The splicing of eukaryotic pre-mRNA is a strictly conserved chemical reaction completed in the nucleus. The process includes: the spliceosome recognizes the exon-intron junction segment, excises the intron through a two-step transesterification reaction and Join adjacent exons. [0003] Although an intron is a sequence in a gene that is only transcribed but not translated, more and more studies...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12N15/82A01H5/00
Inventor 黄大昉郎志宏朱莉潘阳阳
Owner THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI
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