Application of polypeptide and small molecules in regulating protein expression level in rice or wheat
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A protein and fusion protein technology, applied in the biological field
Inactive Publication Date: 2017-01-11
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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[0006] Currently, the improved Shield1-RDDK system has been verified in the dicot model plant Arabidopsis ( figure 1 ), this is the first method to use small molecules to directly regulate protein accumulation levels in plants, but there are no relevant research reports on important food crops such as rice and wheat
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Embodiment 1
[0046] Embodiment 1. System for cultivating functional transgenic crops
[0047] Systems for breeding resistant crops include peptide-related biomaterials and small molecule shield1.
[0048] Peptides and related biological materials are as follows:
[0049] Polypeptide RDDK or RDDKHA, polypeptide UbRDDK or UbRDDKHA, polypeptide encoding gene UbRDDK or UbRDDKHA, expression vector containing polypeptide encoding gene, fusion protein obtained by fusion of polypeptide encoding gene and functional protein gene, and fusion protein encoding gene.
[0050] The amino acid sequence of the RDDK polypeptide is the 77th-185th amino acid from the N-terminal of the sequence 2 in the sequence listing, the coding gene RDDK of the RDDK polypeptide is the 229-555th nucleotide from the 5' end of the sequence 1 in the sequence listing, and the UbRDDK polypeptide The amino acid sequence of the amino acid sequence is sequence 2 in the sequence listing from the 1-185th amino acid at the N-terminal,...
Embodiment 2
[0054] Embodiment 2, the application for cultivating functional transgenic crops
[0055] 1. Polypeptide RDDKHA / RDDK fusion functional protein expression
[0056] 1. Recombinant vector
[0057] The pCambia2300-Ubi vector is a recombinant vector obtained by inserting the maize ubiquitin promoter shown in sequence 3 into the pCambia2300 (from http: / / www.cambia.org / daisy / cambia / 585.html) vector digested by EcoRI, named is pCambia2300-Ubi.
[0058] pCambia2300Ubi-UbRDDK intermediate vector: replace the UbRDDK gene fragment shown at positions 1-555 of Sequence 1 with the DNA fragment between the Sac I and Kpn I double restriction sites of the pCambia2300-Ubi vector backbone to obtain the intermediate vector pCambia2300Ubi-UbRDDK.
[0059] pCambia2300Ubi-UbRDDKHA intermediate vector: replace the UbRDDKHA gene fragment shown in Sequence 1 with the DNA fragment between the Sac I and Kpn I double restriction sites of the pCambia2300-Ubi vector backbone to obtain the intermediate vect...
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Abstract
The invention discloses application of polypeptide and small molecules in regulating a protein expression level in rice or wheat. The invention provides application of a polypeptide related biomaterial and shield1 in cultivating genetically modified crops. The polypeptide related biomaterial is any one of polypeptide RDDK or RDDKHA, a polypeptide RDDK encoding geneor or a polypeptide RDDKHA encoding gene, an expression vector containing the polypeptide RDDK encoding gene or an expression vector containing the polypeptide RDDKHA encoding gene, and a fusion protein and a fusion protein encoding gene formed by fusing the polypeptide RDDK encoding gene or the polypeptide RDDKHA encoding gene and a functional protein gene. The experiments provided by the invention show that an efficient space-time controlled chemical induction system of the transgene protein expression level is built in the rice and the wheat for the first time, and finally a powerful technical support is provided for crop function gene excavation and transgene molecule breeding.
Description
technical field [0001] The invention relates to the field of biotechnology, in particular to a method of using polypeptides and small molecules to regulate protein expression levels in rice or wheat. Background technique [0002] The chemical inducible system of plant gene expression has been developed in various forms after years of research, such as the tetracycline inducible system derived from bacteria, the glucocorticoid inducible system derived from mammals, the copper ion induced system derived from yeast, and the gene expression induced by ethanol control system, etc. Due to the strong controllability of the chemical induction system, it has been widely used in plant biology research. Regrettably, methylcortisol, estradiol, tetracycline and heavy metals used as inducers in the above chemical regulation systems are all harmful to the ecological environment and should not be used in production practice. In addition, there are many other problems in the chemically ind...
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