Cultivation of sheath blight-resisting transgenic paddy rice and special vector
A technology of sheath blight and coding genes, applied in the field of sheath blight-resistant transgenic rice cultivation and special vectors, can solve the problems of poor control effect, untimely application of medicines, incorrect methods, etc.
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Embodiment 1
[0035] Embodiment 1, containing GAFP 2 High-efficiency expression vector pCambia2300-35S-GAFP 2 build
[0036] Synthetic GAFP 2 Gene, the nucleotide sequence of GAFP2 is sequence 1 in the sequence listing, and the amino acid sequence encoded by the gene is sequence 2 in the sequence listing. Sequence 1 in the above sequence list consists of 516 nucleotides, the 1st-84th position from the 5' end is the gene encoding the signal peptide, and the 85th-516th position from the 5' end is the gene encoding GAFP 2 protein gene. Sequence 2 in the above sequence list consists of 171 amino acids, the 1-28th position from the amino terminal is a signal peptide, and the 29-171st position from the amino terminal is GAFP 2 protein.
[0037] GAFP 2 The gene (sequence 1 in the sequence listing) was connected to pUCm-T (purchased from Shanghai Sangon Bioengineering Co., Ltd.), and the vector pUCm-GAFP was constructed 2 .
[0038] Digest pUCm-GAFP with XbaI and SacI 2 , to obtain GAFP af...
Embodiment 2
[0040] Embodiment 2, transfer GAFP 2 Study on the Acquisition and Function of Rice
[0041] 1. Transfer to GAFP 2 harvesting of rice
[0042] 1. Convert to GAFP 2 harvesting of rice
[0043] 1) the expression vector pCambia2300-35S-GAFP obtained in Example 1 2 Transformed into Agrobacterium tumefaciens LBA4404 (purchased from Invitrogen) to obtain Agrobacterium tumefaciens LBA4404 / pCambia2300-35S-GAFP 2 .
[0044] 2) Pick Agrobacterium tumefaciens LBA4404 / pCambia2300-35S-GAFP respectively 2 Single colonies were inoculated in 20ml of YEP medium containing 50mg / L kanamycin, 50mg / L rifampicin and 30mg / L streptomycin, and cultured overnight at 28°C and 280rpm until the OD600 was 0.7-0.8. Then centrifuge at 5000 rpm, discard the supernatant, and resuspend the pellet with NB-liquid medium. Finally, the rice "Xudao 3" (Xu Jun, Duan Xiangmao, Liu Qiang, Yang Bo, Xu Zongjin, Zhou Wei, Chen Liugen and Zhong Weigong. 2008. Experimental Research on Super-high Yield Cultivation of ...
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