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
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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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