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Sequences of Saccharum officinarum sucrose phosphate synthase B (SofSPSB) gene and encoded protein thereof

A technology of phosphate synthase and protein coding, which is applied in the direction of enzymes, enzymes, biochemical equipment and methods, and can solve problems such as lack of research

Inactive Publication Date: 2011-08-17
广西壮族自治区甘蔗研究所
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Problems solved by technology

However, only one complete cDNA sequence of the SPS gene has been cloned in sugarcane so far, and no further studies have been reported

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  • Sequences of Saccharum officinarum sucrose phosphate synthase B (SofSPSB) gene and encoded protein thereof
  • Sequences of Saccharum officinarum sucrose phosphate synthase B (SofSPSB) gene and encoded protein thereof
  • Sequences of Saccharum officinarum sucrose phosphate synthase B (SofSPSB) gene and encoded protein thereof

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specific Embodiment approach

[0017] 1. Plant SPS (sucrose phosphate synthase) gene evolution analysis: use "sucrose phosphate synthase" to search the NCBI website (http: / / www.ncbi.nlm.nih.gov) non-redundant amino acid database to obtain all plant SPS sequences. All sequences obtained were analyzed to remove repetitive sequences. A total of 77 plant SPS sequences were obtained, including 43 full-length sequences. Align the amino acid sequences of all full-length SPS with ClustalX, and save the alignment results in PHYLIP format; then use the Seqboot, Protdist, Neighbor and Consense programs of the PHYLIP software package to perform evolutionary analysis, and divide all full-length SPS genes in plants for the corresponding family.

[0018] 2. Design primers for amplifying the core fragment of the SoSPSB gene: compare the cDNA sequences of the plant B family SPS gene, and design a pair of primers for amplifying the partial fragment of SofSPSB in the conserved region:

[0019] SofSPSB-F(5'-atgaacttcaacccctc...

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Abstract

The invention discloses sequences of a Saccharum officinarum sucrose phosphate synthase B (SofSPSB) gene and an encoded protein thereof. The sequences are obtained by the following steps: carrying out evolution analysis of full-length sequences of sucrose phosphate synthase genes of all plants, designing primers based on the conservative regions of the same family of sucrose phosphate synthase gene sequences, extracting total RNA from Saccharum officinarum tender leaves, carrying out reverse transcription, amplifying with a conventional PCR (polymerase chain reaction) method, and cloning into the full-length cDNA sequence of SofSPSB gene in combination with the RACE (rapid amplification of cDNA ends) technique. The invention not only studies the transcription and expression mechanisms of sucrose phosphate synthase so as to lay a foundation for further discussion of a sucrose accumulation mechanism; and can obtain a purified protein with biological activity through an amino acid sequence so as to provide a basis of studying the biological function of sucrose phosphate synthase and improving the crop variety by using the SofSPS gene.

Description

technical field [0001] The invention relates to a key enzyme gene for controlling sucrose synthesis in a plant and its coded protein sequence. Background technique [0002] Sucrose phosphate synthase (hereinafter referred to as SPS) is a key enzyme that controls sucrose synthesis in higher plants. The accumulation of sucrose in plants is positively correlated with the activity of SPS. In addition, SPS is also involved in the growth and yield formation of plants, and plays an important role in plant stress resistance, Such as cold resistance, drought resistance and salt tolerance play an important role. [0003] The research on the transgenic SPS gene was first carried out on tomato in the early 1990s. When the maize SPS gene was introduced into the tomato, the SPS activity in the transgenic tomato leaves increased by 6 times, the starch content in the leaves decreased and the sucrose content increased. Afterwards, studies on transgenic SPS genes were carried out on various ...

Claims

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Application Information

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IPC IPC(8): C12N15/52C12N9/00
Inventor 李杨瑞黄东亮黎涛田智得李双喜汪淼桂意云廖青梁俊吴凯朝方锋学
Owner 广西壮族自治区甘蔗研究所
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