Acta Veterinaria et Zootechnica Sinica ›› 2025, Vol. 56 ›› Issue (1): 307-318.doi: 10.11843/j.issn.0366-6964.2025.01.028
• Preventive Veterinary Medicine • Previous Articles Next Articles
CAI Yunfeng1,2(
), LI Hong1,2, HUANG Xiaoming1,2, HE Qing1,2, DING Zhenyu1,2, YU Wanting3, LUO Shile4, CHEN Fangzhi5, WANG Naidong1,2, YANG Yi1,2, ZHAN Yang1,2,*()
Received:2024-02-19
Online:2025-01-23
Published:2025-01-18
Contact:
ZHAN Yang
E-mail:Yvonne_2019@stu.hunau.edu.cn;yangzhan@hunau.edu.cn
CLC Number:
CAI Yunfeng, LI Hong, HUANG Xiaoming, HE Qing, DING Zhenyu, YU Wanting, LUO Shile, CHEN Fangzhi, WANG Naidong, YANG Yi, ZHAN Yang. Study on the Presentation Strategy and Immunogenicity of Porcine Parvovirus Epitope Inserted by 3-fold Axes Region on the Surface of Porcine Circovirus Type 2 Virus-like Particles[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(1): 307-318.
Fig. 1
Schematic diagrams of the mutants of the PCV2 Cap The residues of Caps, exposed on the capsid surface, are labelled with open circles on the bottom, other residues by filled circles (using the 3D structure of the PCV2 capsid as a reference[6])"
Table 1
Primer information"
| 引物名称 Primer name | 引物序列(5′→3′) Primer sequence (5′→3′) |
| PCV2-cap-F1 | CGCCATATGCGGGGTTCTCATCATC |
| PCV2-cap-R1 | CGCGGATCCTTATTTCGGATTCAGCGG |
| PCV2-GH-D1B-F2 | TCACCCCGAAACCGTTTCAACCGAACAATAAGC |
| PCV2-GH-D1A-R2 | GTTGAAACGGTTTCGGGGTGAAATAACG |
| PCV2-GH-D2B-F2 | CTGTGGCTGCGCCTGCAACACGTGGGTCTGGGCACC |
| PCV2-GH-D2A-R2 | CCCACGTGTTGCAGGCGCAGCCACAGCTGG |
| PCV2-GH-M1B-F2 | CCG$\underline{{\rm{GCCGCAGCGGCCGCAGCGGCCGCG}}}$TTTCAACCGAACAATAAG |
| PCV2-GH-M1A-R2 | GAAACG$\underline{{\rm{CGGCCGCTGCGGCCGCTGCGGC}}}$CGGTTTCGGGGTGAAA |
| PCV2-GH-M2B-F2 | CAA$\underline{{\rm{GCCGCAGCGGCCGCAGCG}}}$CACGTGGGTCTGGGC |
| PCV2-GH-M2A-R2 | GTG$\underline{{\rm{CGCTGCGGCCGCTGCGGCTTG}}}$CAGGCGCAGCCA |
| PCV2-GH-M3B-F2 | CCG$\underline{{\rm{CGCCGTCGCCGTCGCCGTCGCCG}}}$TTTTCAACCGAACAAT |
| PCV2-GH-M3A-R2 | GAAA$\underline{{\rm{ACGGCGACGGCGACGGCGACGGCG}}}$CGGTTTCGGGGTGAA |
| PCV2-GH-M4B-F2 | CAAC$\underline{{\rm{GCCGTCGCCGTCGCCGTC}}}$ACGTGGGTCTGGGC |
| PCV2-GH-M4A-R2 | GTG$\underline{{\rm{ACGGCGACGGCGACGGCG}}}$TTGCAGGCGCAGCCA |
| PCV2d-GH-I1B-F2 | AACCG$\underline{{\rm{CAGCAGATTACCGATGCG}}}$TTTCAACCGAACAAT |
| PCV2d-GH-I1A-R2 | GAAA$\underline{{\rm{CGCATCGGTAATCTGCTG}}}$CGGTTTCGGGGTGAAATAAC |
| PCV2d-GH-I2B-F2 | CTGCAACAG$\underline{{\rm{CAGATTACCGATGCGCACGTGGGTCTGGG}}}$CACC |
| PCV2d-GH-I2A-R2 | CACGTG$\underline{{\rm{CGCATCGGTAATCTGCTG}}}$TTGCAGGCGCAGCCAC |
Fig. 2
Distribution characteristics of amino acid residues on the surface of the icosahedral threefold axes of PCV2 capsid and simulation of exogenous epitopes"
Fig. 3
Identification of recombinant plasmids by digestion M. DL5000 DNA marker; 1. The digested product of PCV2-Cap; 2-5. The digested products of GH-D1, GH-M1, GH-M3, GH-I1; 6-9. The digested products of GH-D2, GH-M2, GH-M4, GH-I2"
Fig. 4
SDS-PAGE analysis of mutant protein (Loop GH) M. Protein marker; T. Total protein after IPTG induction; S. Supernatant after ultrasonic cracking process; P. Precipitation after ultrasonic cracking process"
Fig. 5
SDS-PAGE (A) and Western blot (B & C) analysis of chimeric epitope recombinant protein (Loop GH) M. Protein marker; 1. Total protein after IPTG induction (GH-I1); 2. Supernatant after ultrasonic cracking process (GH-I1); 3. Precipitation after ultrasonic cracking process (GH-I1); 4. Total protein after IPTG induction (GH-I2); 5. Supernatant after ultrasonic cracking process (GH-I2); 6. Precipitation after ultrasonic cracking process (GH-I2)"
Fig. 6
Purification of Loop GH related mutant protein M. Protein marker; 1. Supernatant after ultrasonic cracking process; 2. Flow-through (Proteins not bound to columns); 3. Proteins that are washed; 4-6. Elution of the target protein"
Fig. 7
PCV2 VLPs and cVLPs were subject to examination by TEM and gel filtration chromatography A. BSA(Control, 66.43 ku); B. PCV2 VLPs(VLPs-WT); C. cVLPs (GH-I1). Bar=100 nm"
Fig. 8
Entry of PCV2 VLPs and cVLPs into PK15 cells identified by indirect immunofluorescence assay Bar=50 μm"
Fig. 9
Evaluation of the immunogenicity of Loop GH displaying exogenous antigen epitopes A. ELISA detection of PCV2 VLPs antibodies; B. ELISA detection of B5-E1; C. SDS-PAGE for PPV VP2; D. Western blot for PPV VP2; E. ELISA detection of PPV VP2. **.P < 0.01"
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