Expression vector for improving content and quality of protein in rice corn and its making method and use

A protein content and expression carrier technology, applied in the field of plant genetic engineering and genetic engineering, can solve the problems of insignificant increase in protein content, low increase in protein quality, that is, the content of limiting amino acids, and avoid the safety of genetically engineered food Effect

Inactive Publication Date: 2007-05-23
SHANGHAI NORMAL UNIVERSITY
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  • Abstract
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  • Claims
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AI Technical Summary

Problems solved by technology

[0012] The technical problem to be solved by the present invention is to provide an expression carrier that utilizes the glycinin gene to improve the protein content and quality of rice and its preparation method and application, so as to overcome the existing expression vector that utilizes the glycinin gene to improve the protein content and quality of rice. Both are composed of rice glutenin GluB-1 and Gt1 gene promoters and soybean Gy3 gene coding sequence cDNA to form a fusion gene, the protein content of the obtained transformants and their offspring rice is not significantly improved, and the quality of the protein is the improvement of the limiting amino acid content. low magnitude defects

Method used

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  • Expression vector for improving content and quality of protein in rice corn and its making method and use
  • Expression vector for improving content and quality of protein in rice corn and its making method and use

Examples

Experimental program
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Embodiment 1

[0048] This example is to illustrate the method of constructing the Gy7 glycinin gene expression vector guided by the Gt1 gene promoter and the structural composition of the obtained vector.

[0049] Methods such as conventional PCR amplification, restriction endonuclease digestion and ligation involved in the operation of constructing the expression vector mainly refer to "Molecular Cloning Experiment Guide" (Sambrook J et al., Molecular Cloning Experiment Guide (Second Edition), Science Press, 1992). For the experimental methods that do not specify specific conditions in the examples, usually follow the conventional conditions or the conditions suggested by the manufacturer.

[0050] The construction process of the two expression vectors of the high-quality glycinin gene guided by the Gt1 gene promoter is as follows:

[0051] The rice glutelin gene Gt1 promoter sequence published in Genbank AY649098 was compared with NCBI to obtain the Genbank AC021891 gene sequence with 99...

Embodiment 2

[0056]This example is to illustrate how to use the expression vector constructed in Example 1 to transform rice.

[0057] Using the above two expression vectors and the pCAMBIA1301 vector containing the resistance selection marker gene and the reporter gene to co-transform the rice varieties with a protein content of 7%-9% in the current conventional rice, the root cancer involved in the cultivation of transgenic rice Agrobacterium culture and resistant callus selection and plant regeneration are mainly carried out according to the method of Liu Qiaoquan (Liu Qiaoquan et al., Acta Physiological Plant, 1998, 24(3): 259-271), and the genetic transformation of rice mainly refers to Li Jianyue et al. (Li Jianyue et al., Science Bulletin, 2004, 49(24): 2556-2561), that is, transgenic offspring without resistance selection marker genes and reporter genes can be obtained, and the protein content and quality of rice can be improved.

[0058] In this example, the Gt1 promoter with a hi...

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Abstract

The invention discloses an expression vector for increasing rice protein content and quality and its production method and application, having the structure and characteristics as a and b shown. The vector contains glutelin gene promoter, soybean Gy7 full length gene or Gy7 gene coding sequence cDNA fragment, terminator and incision enzyme sensitive sequence or marker gene sequence. The vector can be used for transforming Gramineae plants like rice to obtain rice variety containing high protein content and high restriction amino acid content.

Description

technical field [0001] The invention belongs to the field of genetic engineering, more specifically relates to the field of plant genetic engineering. Background technique [0002] The nutritional components of rice include starch, protein, fat, vitamins, etc., while the nutritional quality of rice mainly refers to the content of protein and essential amino acids. At present, the total protein content of conventional rice cultivars is relatively low, generally 7%-9%. Zhang Ruipin and others believe that in areas where rice is the staple food, polished rice provides at least 40% of protein. If the protein content of rice in our country is increased by 1%-2% on the current basis, it will undoubtedly improve the nutritional level of our people. Great contribution (Zhang Ruipin et al., Rice Quality Breeding, Crop Quality Breeding (Edited by Liu Houli), Hubei Science and Technology Press, 2001, pp: 37). Rice high-protein breeding is more important for human health, especially f...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82C12N15/12
Inventor 李建粤顾婷玉米东李海权石少华
Owner SHANGHAI NORMAL UNIVERSITY
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