cxcl13 DNA vaccine and its application
A DNA vaccine and vaccine technology, applied in the field of medicine, can solve the problems of human body toxicity and side effects, and achieve the effect of high potency
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Embodiment 1
[0023] Example 1: Construction and identification of pcDNA3.1-HBc-mCXCL13
[0024] 1. Construction of pcDNA3.1-HBc-mCXCL13
[0025] In order not to affect the original spatial structure of CXCL13 as much as possible so that it can present the original epitope, HBc and CXCL13 were used figure 1 and 2 The indicated linker was connected, and the In-Fusion cloning technique was used to connect to the vector pcDNA3.1(-).
[0026] Cloning primers are as follows:
[0027] 0-linker:
[0028] HBc-HF:
[0029] TGCTGGATATCTGCAGAATTCACGTCATGGACATCGACCCTTAAAG
[0030] HBc-1: TGTGCTGAGCCTCATTGCTGGATCTTCCAAATTAACAC
[0031] mCXCL13-F: ATGAGGCTCAGCACAGCAAC
[0032] mCXCL13-R: TACTAGGTCTCTAGA GGCAGCTCTTCTCTTACTCAC
[0033] HBc-2: TCTAGAGACCTAGTAGTCAGTT
[0034] HBc-TF: CAGCGGTTTAAACTTAAGCTTCTAACATTGAGGTTCCCGAGAT
[0035] S-linker:
[0036] HBc-HF:
[0037] TGCTGGATATCTGCAGAATTCACGTCATGGACATCGACCCTTAAAG
[0038] HBc-1: ACCGCTACCACCACCTGCTGGATCTTCCAAATTAACAC
[0039] mCXCL13-F: GGT...
Embodiment 2
[0057] Example 2: Expression identification of pcDNA3.1-HBc-mCXCL13 in HEK 293T
[0058] After the plasmid was transfected into HEK 293T cells, the expression of HBc-mCxcl13 was successfully detected by RT-PCR and Western Blot.
Embodiment 3
[0059] Example 3: Antibody titers in serum after CXCL13 vaccination are high
[0060] Orbital venous blood was collected, and serum was separated. Enzyme-linked immunosorbent assay (ELLSA) was used to detect high anti-CXCL13 antibody titers at 1 month, 2 months, 4 months and 8 months after immunization.
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