Type 31 recombinant human papillomavirus virus-like particle and its preparation method
A human papillomavirus and fusion gene technology, which is applied in the field of 31 recombinant human papillomavirus virus-like particles and its preparation, can solve the problems of difficult large-scale production, large protein loss, and uneven particles
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment l
[0070] Embodiment 1: the design and synthesis of the HPV L1 gene of codon optimization
[0071] The gene sequences were derived from the various types of HPV sequences published on PUBMED. All HPV DNA sequences were synthesized after codon optimization of the selected HPV DNA sequences according to the codon preference of Escherichia coli for gene transcription. Primers were designed according to the synthetic DNA sequence, and the synthetic gene was used as a template for PCR amplification. The resulting codon-optimized sequences were verified by DNA sequencing.
[0072] DNA sequences of various types of HPV before and after optimization:
[0073] SEQ NO.1: DNA sequence of HPV31 type L1 before optimization
[0074] SEQ NO.2: DNA sequence of optimized HPV31 type L1
Embodiment 2
[0075] Example 2: Construction and identification of recombinant vector pGEX-6P-1-GST-HPV31 L1:
[0076] Primers for amplifying the DNA fragment of HPV31 L1: (the restriction sites are BamHI and XhoI, respectively)
[0077] Forward-HPV31 L1-ApaI:5'ACTTCAGGATCC ATGTCTCTGTGGCGTCCGTCTG
[0078] Reverse-HPV31 L1-XhoI:5'ATCTCACTCGAGCTATTTTTTGGTTTTTTTACGTTT
[0079] PCR amplification reaction system: 10 x Pfu buffer 20 μL, Pfu enzyme 4 μL, 10 mM dNTP 2.5 μL, 5’ Primer (5 μM) 10 μL, 3’ Primer (5 μM) 10 μL, template DNA 50 ng, add d2H2O to 200 μL.
[0080] Gene PCR amplification conditions: 95°C for 3 min; 95°C for 30 sec, 58°C for 30 sec, 72°C for 4 min; cycle 32 times; 72°C for 10 min.
[0081] The L1 gene fragment containing BamH I and XhoI restriction sites and the vector pGEX-6P-1 were subjected to BamH I / XhoI double digestion treatment, and then the recovered gene fragment was combined with pGEX-6P-1 containing the corresponding cohesive end using T4 DNA ligase. 6P-1 was subj...
Embodiment 3
[0084] Example 3: Construction of recombinant vector pGEX-6P-1m-GST-SUMO-HPV31 L1 vector
[0085] Construction of pGEX-6p-1m vector: In order to make the ApaI restriction site (GGGCCC) near the multi-restriction site the only ApaI restriction site of the vector, point mutation was carried out without changing the protein expression sequence of the lacI gene ApaI (3890) can be eliminated by changing the Gly codon GGC in another ApaI recognition sequence GGGCCC of the commercially available pGEX-6p-1 vector to its synonymous codon GGT. Through such modification, ApaI can be used to insert and express genes.
[0086] Primers for amplifying the DNA fragment of SUMO: (restriction sites are ApaI and BamHI respectively)
[0087] Forward -SUMO-ApaI: ACTTCAGGGCCCTCTGACCAGGAAGCTAAACCGTC
[0088] Reverse-SUMO-BamHI: CGCGGATCCACCGGTCTGTTCCTGGTAAAC
[0089] Primers for amplifying the DNA fragment of HPV31 L1: (the restriction sites are BamHI and XhoI, respectively)
[0090] Forward-HPV...
PUM
Property | Measurement | Unit |
---|---|---|
diameter | aaaaa | aaaaa |
diameter | aaaaa | aaaaa |
diameter | aaaaa | aaaaa |
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com