Method of increasing lysine content in paddy rice seed and special carrier

A technology of rice seeds and lysine, applied in the direction of introducing foreign genetic material using vectors, botany equipment and methods, biochemical equipment and methods, etc., can solve the problem of not being able to obtain free lysine

Inactive Publication Date: 2006-09-20
BEIJING KAITUO DNA BIOTECH RES CENT
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Problems solved by technology

The limitation of the above-mentioned method for increasing plant lysine content is that free lysine cannot be obtained, and free amino acids are easily absorbed by the human body

Method used

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  • Method of increasing lysine content in paddy rice seed and special carrier
  • Method of increasing lysine content in paddy rice seed and special carrier
  • Method of increasing lysine content in paddy rice seed and special carrier

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Embodiment 1, for improving the construction of the special-purpose plant transformation vector of lysine content in rice seed

[0040] Now refer to figure 2 , to construct a special plant transformation vector for increasing the lysine content in rice seeds, the specific process includes the following steps:

[0041] 1. Construction of the plant transformation vector of the key enzyme gene of lysine synthesis - dihydropicolinate synthase gene DHPS

[0042] A pair of primers were designed according to the cDNA sequence (GenBank number: sequence 2 in the M12844 sequence listing) of the Escherichia coli dihydropyridine carboxylate synthase gene DHPS, and the primer sequences were as follows:

[0043] P1 (upstream primer): 5'-CACCCCAGGCGACTGTCTTCAATATTAC-3

[0044] P2 (downstream primer): 5'-GGCGCGACTTTTGAACAGAGTA-3'

[0045] The genomic DNA of Escherichia coli DH5α was extracted and used as a template. Under the guidance of primers P1 and P2, the cDNA sequence of the ...

Embodiment 2

[0051] Embodiment 2, the acquisition and detection of transgenic rice

[0052] 1. Detection of expression of rice LKR / SDH in rice tissues

[0053] It is reported that LKR / SDH of most plants is specifically expressed in seeds, and its expression is activated by the increase of lysine content, so that the lysine content in plant seeds reaches a balance. In order to verify the expression characteristics of the rice homologous gene (named OsLKR / SDH) of the LKR / SDH obtained in Example 1, the total RNA of the wild-type rice immature embryo, seed, callus, root, flower and leaf is now extracted respectively, to The cDNA obtained by reverse transcription was used as a template, and semi-quantitative RT-PCR detection was carried out under the guidance of primers P3 and P4, and Actin was used as a reference. The detection results were as follows: Figure 4 As shown, the gene is highly expressed in seeds and callus of wild-type rice, slightly expressed in leaves and flowers, and slightly...

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Abstract

This invention discloses method and carrier for improving the content of lysine in rice seeds. The carrier is a plant expression carrier containing a rice seed specific promoter, dihydropicolinic acid synthetase gene and rice lysine-oxoglutaric acid reductases / saccharopin dehydrogenase gene interfering RNA coding sequence. The said dihydropicolinic acid synthetase gene is located at the downstream of the rice seed specific promoter, and the said rice lysine-oxoglutaric acid reductases / saccharopin dehydrogenase gene interfering RNA coding sequence is located at the downstream of the constitutive promoter. The genetically modified rice strains obtained in this invention have much higher free lysine content in seeds (10 times higher than that of Wild rice). Besides, the exogenous gene has no effect on rice growth. The method and the carrier have important applications in improving the lysine content in rice seeds, and provide a new way for modifying rice and other plants.

Description

technical field [0001] The invention relates to a method for increasing plant lysine content and a special carrier thereof, in particular to a method for increasing lysine content in rice seeds and a special carrier thereof. Background technique [0002] Essential amino acids refer to amino acids that cannot be synthesized by the human body or cannot be synthesized at a rate that meets the needs of the human body, and must be ingested from food. Lysine is one of the eight essential amino acids, known as the "first essential amino acid", and is an important component of protein. Lysine plays an important role in the metabolism of organisms, and lysine deficiency can cause protein dysfunction and affect the growth and development of organisms. Lysine has optical activity, and organisms can only use L-form (L-form) lysine. [0003] The Chinese diet structure is mainly based on grains, and the protein intake mainly comes from grains. However, the lysine content in vegetable p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82C12N15/52C12N15/53
Inventor 王喜萍于鲲陈丽娜谢莹夏勉邓兴旺
Owner BEIJING KAITUO DNA BIOTECH RES CENT
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