A kind of recombinant retinol binding protein 4 and its preparation method and application
A technology that binds proteins and retinol, applied in the biological field, can solve problems such as weak immunogenicity of RBP4, poor antibody performance, and restrictions on the application of RBP4
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0030] [Example 1] Preparation of recombinant RBP4(20)
[0031] (1) Recombinant RBP4(20) protein design:
[0032] According to the amino acid sequence of the RBP4 protein published on the NCBI website and the characteristics of the sequence, amino acids 1-18 are signal peptides, and the mature protein sequence is amino acids 19-201, with a total of 183 amino acids. The amino acid sequence is shown in the sequence table SEQID NO. As shown in 4, replace the F at position 20 with Y to obtain the amino acid sequence shown in the sequence listing SEQ ID NO.1.
[0033] (2) Optimization and acquisition of recombinant RBP4(20) gene sequence:
[0034] The mutated amino acid sequence SEQ ID NO.1 was reverse-translated into the nucleotide sequence SEQ ID NO.5 with Escherichia coli dominant codons, and the recombinant RBP4 composed of Escherichia coli dominant codons ( 20) Genes.
[0035] The RBP4(20) gene is designed into two segments A and B. The first round of PCR reaction system is...
Embodiment 2
[0053] [Example 2] Preparation of recombinant RBP4(35)
[0054] (1) Recombinant RBP4(35) protein design:
[0055] In the amino acid sequence of retinol binding protein 4, L at position 35 was replaced with G to obtain the amino acid sequence shown in SEQ ID NO.2 of the sequence table.
[0056] (2) Optimization and acquisition of recombinant RBP4(35) gene sequence:
[0057] The mutated amino acid sequence SEQ ID NO.2 was reverse-translated into the nucleotide sequence SEQ ID NO.6 with the dominant codons of Escherichia coli, and the recombinant RBP4(35) composed of the dominant codons of Escherichia coli was obtained by annealing extension PCR technique Gene.
[0058] The RBP4(35) gene was designed into two segments A and B. The first round of PCR reaction system was 2×PCR reaction solution, double distilled water and primers from Biotec Company. The PCR reaction conditions were 94°C for 1 minute, followed by 94°C for 1 minute , 55°C for 1 minute, 72°C for 1 minute, 5 cycles...
Embodiment 3
[0076] [Example 3] Preparation of recombinant RBP4 (20-35)
[0077] (1) Recombinant RBP4(20-35) protein design:
[0078] In the amino acid sequence of retinol binding protein 4, F at position 20 is replaced by Y while L at position 35 is replaced with G to obtain the amino acid sequence shown in the sequence table SEQ ID NO.3.
[0079] (2) Optimization and acquisition of recombinant RBP4(20-35) gene sequence:
[0080] The mutated amino acid sequence SEQ ID NO.3 was reverse-translated into the nucleotide sequence SEQ ID NO.7 with the dominant codons of Escherichia coli, and the recombinant RBP4 (20- 35) Genes.
[0081] Design the RBP4(20-35) gene into two segments, A and B. The first round of PCR reaction system is 2×PCR reaction solution, double distilled water, and primers from Biotech Company. The PCR reaction conditions are 94°C for 1 minute, and then 1 minute, 55°C for 1 minute, 72°C for 1 minute, 5 cycles; the PCR reaction system for the next two rounds is 2×PCR reacti...
PUM
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