Method for improving expression level of target protein in transgenic rice endosperm

A technology for transgenic rice and target protein, which is applied to the signal peptide coding sequence of glutenin GluA-2 to increase the expression of the target protein in the endosperm of transgenic rice, and can solve the problems of low investment, low expression of the target protein, and high output. problems, to achieve the effect of scientific, simple and advanced methods, and improved expression

Inactive Publication Date: 2011-01-12
YANGZHOU UNIV
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Problems solved by technology

Especially in recent years, using the cheap production system of plants as a bioreactor to mass-produce recombinant heterologous proteins with important functions, medically active polypeptides or vaccines, antibodies, industrial enzymes or proteins, etc., is the current plant gene One of the important fields of engineering development, its research and development are in the ascendant, and it is possi

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  • Method for improving expression level of target protein in transgenic rice endosperm
  • Method for improving expression level of target protein in transgenic rice endosperm
  • Method for improving expression level of target protein in transgenic rice endosperm

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Embodiment Construction

[0015] The specific embodiment of the present invention is further illustrated in four optimal steps below. The contents described in the embodiments do not limit the scope of the claims of the present invention.

[0016] 1. Cloning of rice glutelin GluA-2 signal peptide coding sequence

[0017] The N-terminal of the rice glutelin GluA-2 precursor contains a signal peptide sequence consisting of 24 amino acids. According to the published nucleotide sequence of rice glutelin GluA-2 gene (GenBank Y00687), a pair of primers Gt1P was designed 6 : 5'CA GGATCCA ATGGCATCCATAAATCGCCCC-3' (SEQ ID NO.3) and Gt1P 10 : 5'-CA CCATG GCTAGGGAGCCATCGCACAA-3' (SEQ ID NO.4), respectively add BamHI and NcoI enzyme cutting sites at the 5' end of the primer. The genomic DNA of rice variety Wuyunjing No. 8 was used as a template for PCR amplification. The PCR amplification conditions were 95°C, 5min; 95°C, 50sec, 55°C, 50sec, 72°C, 30sec, 30 cycles; 72°C, 5min . The PCR product was cloned...

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Abstract

The invention discloses a method for improving expression level of a target protein in transgenic rice endosperm by use of a signal peptide coding sequence of a rice storage protein. The method comprises the following steps: connecting a rice GluA-2 gene signal peptide coding sequence with a target protein coding sequence to form a fusion protein gene; transforming the fusion protein gene into rice cells to obtain transgenic rice; and culturing and ripening the transgenic rice, and then expressing the target recombinant protein in seed endosperm. In the invention, the target protein coding sequence and the GluA-2 signal peptide coding sequence containing an SEQ ID NO. 1 sequence are constructed into the fusion gene and the fusion gene is led into a plant, which can obviously improve the expression level of the target protein in the transgenic rice endosperm.

Description

technical field [0001] The invention relates to a method for increasing the expression level of a target protein in transgenic rice endosperm. More specifically, the present invention relates to the signal peptide coding sequence of glutelin GluA-2, one of the rice seed storage proteins, and the research and application of the sequence. Fusion of the sequence with the target protein can significantly improve the efficiency of the target protein in transgenic Expression levels in rice endosperm. The invention belongs to the field of biological genetic engineering. Background technique [0002] Using transgenic technology to express heterologous genes in plants to improve stress resistance, increase yield and improve quality has become the focus and focus of modern agricultural biotechnology research and development. Especially in recent years, using the cheap production system of plants as a bioreactor to mass-produce recombinant heterologous proteins with important functio...

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Application Information

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IPC IPC(8): C12N15/67C12N15/62A01H5/00
Inventor 刘巧泉顾铭洪
Owner YANGZHOU UNIV
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