Late blight resistance-related proteins and their related biomaterials and applications
A biomaterial and protein technology, applied in the field of late blight resistance-related proteins and related biomaterials, can solve the problems of insensitivity to fungicides, high cost, human health and environmental safety impacts, etc.
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[0063] Embodiment 1, the cloning of SP-LBR3 gene and its functional verification
[0064] 1. Cloning of SP-LBR3 gene
[0065] Genomic DNA of tomato (Solanum lycopersicum) CLN2037B was used as a template, and primers SP-LBR3EF3 (5'-taacctgcaggTTCAAACCATCTTCATAGAGGC-3') and SP-LBR3ER3 (5'-attggcgcgccTGGGGCTTAGAAAAAGGTTG-3') were obtained by using Phusion high-fidelity DNA polymerase (Thermo Fisher, Waltham, MA, USA) PCR amplified the fragment of 4kb upstream of SP-LBR3 gene, SP-LBR3 gene and downstream 2kb of SP-LBR3 gene, and this fragment was called SP-LBR3 gene expression cassette PCR product. The PCR product of the SP-LBR3 gene expression cassette was cloned into pCR-Blunt II-TOPO (Invitrogen, Carlsbad, CA, USA) to obtain a recombinant vector pCR-SP-LBR3 containing the PCR product of the SP-LBR3 gene expression cassette. Sequencing results show that the DNA sequence of the PCR product of the SP-LBR3 gene expression cassette is as SEQ ID No.1, which consists of 8007 nucleoti...
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