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Sugarcane neutral/alkaline invertase gene and encoded protein sequence thereof

A protein sequence and invertase technology, applied in the field of bioengineering, can solve problems such as cloning

Inactive Publication Date: 2013-01-23
GUANGXI UNIV
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0005] At present, there are many studies on acid invertase, but less research on neutral / alkaline invertase, and only a few kinds of plants have cloned the full-length sequence of neutral / alkaline invertase gene. No one has cloned the gene in sugarcane

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  • Sugarcane neutral/alkaline invertase gene and encoded protein sequence thereof
  • Sugarcane neutral/alkaline invertase gene and encoded protein sequence thereof

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Embodiment Construction

[0029] Implementation example

[0030] 1. Evolutionary analysis of plant alkaline / neutral invertase genes:

[0031] Use "alkaline / neutral invertase" to search the NCBI website (http: / / www.ncbi.nlm.nih.gov) non-redundant amino acid database to obtain all plant neutral / alkaline invertase gene sequences. All the obtained sequences were analyzed, and repeated sequences were removed, and a total of 20 plant neutral / alkaline invertase sequences were obtained, including 10 full-length sequences. Align the amino acid sequences of all full-length neutral / alkaline invertases with ClustalX, and save the alignment results in PHYLIP format; then use the Seqboot, Protdist, Neighbor and Consense programs of the PHYLIP software package for evolutionary analysis, and plant All full-length neutral / alkaline invertase genes are grouped into corresponding families.

[0032] 2. Design primers for amplifying the core fragment of the neutral / alkaline invertase gene:

[0033] Compare the cDNA seque...

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Abstract

The invention discloses a sugarcane neutral / alkaline invertase gene and an encoded protein sequence thereof. On the basis that all plant neutral / alkaline invertase gene full-length sequences are subjected to evolution analysis, primers are designed in a conserved region of neutral / alkaline invertase gene sequences of the same family, total ribose nucleic acid (RNA) is extracted from tender leaves of sugarcane, the total RNA is subjected to inverse transcription and then is amplified by the conventional polymerase chain reaction (PCR) method, then the amplified total RNA is cloned by a rapid amplification of complementary deoxyribonucleic acid (cDNA) ends (RACE) technology so as to obtain the full-length cDNA sequences of the neutral / alkaline invertase gene; and through VectorNTI software analysis, the open reading frame (ORF) of the obtained sequence is 1,812bp, and 603 amino acids are encoded. By the gene and the method, basis is founded for research on the transcription and expression mechanism of neutral / alkaline invertase and further discussion of accumulation mechanisms of sugarcane; and purified protein with the biological activity can be obtained by the amino acid sequence, and basis is provided for researching the biological function of the neutral / alkaline invertase.

Description

technical field [0001] The invention belongs to the technical field of bioengineering and relates to a sugarcane neutral / alkaline invertase gene and its encoded protein sequence. Background technique [0002] There are many isozyme forms of invertase in tissues of higher plants. According to the optimum pH, they can be divided into two categories: acid invertase and neutral / alkaline invertase. Alkaline / Neutral Invertase (Alkaline / Neutral Invertase) is a cytoplasmic enzyme with an optimum pH value of around 7.0-8.0. The enzyme only uses sucrose as a substrate, and can irreversibly decompose sucrose into glucose and fructose for energy metabolism. In plant tissues with low acid invertase and sucrose synthase activities, neutral / alkaline invertase breaks down sucrose and provides a substrate for the Krebs cycle, so it is also considered a "maintenance enzyme". [0003] At present, genes of such enzymes have been cloned in plants such as wheat, corn, rice, cassava, and small ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/52C12N9/00
Inventor 王爱勤牛俊奇何龙飞杨丽涛李杨瑞
Owner GUANGXI UNIV