Expression and purification of recombinant coat protein of sugarcane mosaic virus from Indonesian isolate as an antigen for antibody production

Expression and purification of recombinant coat protein of sugarcane mosaic virus from Indonesian isolate as an antigen for antibody production

Natalia Tri Astuti;Nurmalasari Darsono;Suvia Widyaningrum;Widhi Dyah Sawitri;Sri Puji Astuti;Win Darmanto;
indonesian journal of biotechnology 2019
331
astuti2019expressionindonesian

Abstract

Sugarcane mosaic virus (SCMV, genus Potyvirus , familia Potyviridae ) is a prominent pathogen of sugarcane ( Saccharum sp. hybrids ). It can cause losses in susceptible varieties, in crop as well as sugar production economically. Although it has been studied in major sugar production countries, researches on the definement of SCMV from Indonesian isolate based on molecular study are very limited. This study aimed to obtain a proper recombinant antigens emanating from coat protein of SCMV from Indonesian isolate for polyclonal antibody production that can be used for immunodiagnosis purposes. A gene encodes the coat protein of SCMV (CP-SCMV) was amplified by the reverse transcription polymerase chain reaction and cloned into vector pJET1.2. The cDNA was inserted into 6X His-tag expression plasmid of pET28a(+) and overexpressed in Escherichia coli BL21(DE3) to produce recombinant protein. The highest expression had performed on 0.1M IPTG induction media for 5 hours at 37 o C. SDS-PAGE analysis clarified that recombinant CP-SCMV remains as an insoluble fraction. The purifications were performed by the affinity Ni-NTA resin which followed by electroelution to obtain highly purified proteins. To meet quality requirements of a proper antigen the highly purified protein was concentrated. Molecular weight of rCP-SCMV (approximately 40 kDa) was clearly observed by 10% SDS-PAGE at the concentration of 16.184 mg/mL. 

Citation

ID: 33862
Ref Key: astuti2019expressionindonesian
Use this key to autocite in SciMatic or Thesis Manager

References

Blockchain Verification

Account:
NFT Contract Address:
0x95644003c57E6F55A65596E3D9Eac6813e3566dA
Article ID:
33862
Unique Identifier:
10.22146/ijbiotech.45551
Network:
Scimatic Chain (ID: 481)
Loading...
Blockchain Readiness Checklist
Authors
Abstract
Journal Name
Year
Title
5/5
Creates 1,000,000 NFT tokens for this article
Token Features:
  • ERC-1155 Standard NFT
  • 1 Million Supply per Article
  • Transferable via MetaMask
  • Permanent Blockchain Record
Blockchain QR Code
Scan with Saymatik Web3.0 Wallet

Saymatik Web3.0 Wallet