siRNA recombinant slow virus for inhibiting human hephaestin gene expression and application
A ferroportin and recombinant lentivirus technology, applied in the fields of molecular biology, biomedicine and genetic engineering, to achieve the effects of regulating metabolism and absorption, regulating serum iron levels, and treating iron metabolism-related diseases
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[0042] According to the human ferroportin auxiliary protein gene (HEPH) sequence (NM_000617.2), the template strand and coding strand were designed. Add BglII and XhoI restriction sites to the 5' end respectively, and send the sequence to Invitrogen Company for synthesis. After screening, the HEPH interference fragment sequence of the present invention with the most significant interference effect is obtained as follows: preferred embodiment
[0043] Template strand (Seq ID No.2):
[0044] 5’-GATCCCCAGGGGTGGATAAAGAATTCTTCAAGAGAGAATTCTTTATCCA
[0045] CCCCTTTTTTTA-3'
[0046] Coding chain (Seq ID No.3):
[0047] 5’-AGCTTAAAAA AGGGGTGGATAAAGAATTC TCTCTTGA
[0048] AATTCTTATCCACCCCT GGGG-3'
[0049] Retro-HEPHi system construction
[0050] The synthesized HEPH interference fragments (Seq ID No.2 and Seq ID No.3) were annealed. Specific steps are as follows:
[0051] 1. Establish an annealing system
[0052] 5ul 10x annealing buffer
[0053] 2nmole template chain Seq ID ...
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