Human bace1 gene transcription promoter and its main cis-acting element and application
A technology of gene transcription and promoter, which can be used in recombinant DNA technology, DNA/RNA fragments, medical preparations containing active ingredients, etc., and can solve very few problems.
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[0025] 1. Materials and methods
[0026] 1.1 Search the GenBank database and literature to determine the promoter region of the BACE1 gene.
[0027] 1.2 Deletion plasmid construction
[0028] The BACE1 promoter luciferase reporter gene p3BU-1273 / -400 contains the sequence from -1273 to -400 upstream of the translation initiation site of the BACE1 gene. The target fragment is amplified by PCR and named as B1U-1273 / -400 or B1U1273-400 (the following names are all the same as this method, B1U-1273 / -400 represents the sequence from the 1273rd nucleotide to the 400th nucleotide upstream of the translation initiation site of BACE1 gene), the primer is B1U1273F / B1U400R (see Table 1), the template is bacterial artificial chromosome DNA (RP11-970G21) (from BAC DNA library). The LA TAQ enzyme from TAKARA company was used for amplification. The reaction program was initial denaturation at 95°C for 30s, followed by 30 cycles at 98°C for 30s, 68°C for 2min, and extension at 72°C for 10mi...
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