Increasing plant growth and yield by using a phenylalanine ammonia lyase sequence
一种核酸序列、作物产量的技术,应用在使用载体引入外来遗传物质、植物肽、生物化学设备和方法等方向,能够解决技术费力等问题
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Embodiment 1
[0105] Example 1 - Construction of PAL Plant Transformation Vectors
[0106] An open reading frame encoding the maize PAL protein was synthesized. The open reading frame comprises SEQ ID NO:1, which encodes the protein sequence of SEQ ID NO:2. Appropriate restriction sites are included at the 5' and 3' ends of the coding sequence to permit cloning of the DNA into plant transformation vectors containing genetic elements suitable to control gene expression. In each plant transformation construct, the PAL open reading frame was located downstream of the plant promoter and 5' untranslated region (5'UTR) and upstream of the 3'UTR. Table 2 summarizes the constructed plant transformation constructs containing the PAL open reading frame.
[0107] Table 2: PAL plant transformation constructs
[0108]
[0109] In addition to the monogenic PAL plant transformation constructs listed in Table 2, multigenic plant transformation constructs containing the PAL gene cassette and the secon...
Embodiment 3
[0117] Example 3 - Transformation of Zea mays
[0118] A. tumefaciens cells carrying the PAL plant transformation vector were used to transform maize (Zea mays cv. B104) cells suitable for regeneration on tissue culture medium. After transformation of maize cells with relevant plant transformation vectors and regeneration of maize plants, PCR analysis was performed to confirm the presence of the gene of interest in the maize genome. Transformation vector number 131556 was used for maize transformation.
Embodiment 4
[0119] Example 4 - Transformation of Rice (Oryza sativa)
[0120]A. tumefaciens cells carrying the PAL plant transformation vector were used to transform rice (Oryza sativa cv. Kitaake) cells suitable for regeneration on tissue culture medium. After transforming rice cells with relevant plant transformation vectors and regenerating rice plants, PCR analysis was performed to confirm the presence of the gene of interest in the rice genome. Transformation vector number 130925 was used for rice transformation resulting in 15 PCR-validated transgenic plants containing the PAL gene cassette.
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