TY - JOUR
T1 - Expression, purification, and characterization of biologically active full-length Mason-Pfizer monkey virus (MPMV) Pr78Gag
AU - Pitchai, Fathima Nuzra Nagoor
AU - Ali, Lizna
AU - Pillai, Vineeta Narayana
AU - Chameettachal, Akhil
AU - Ashraf, Syed Salman
AU - Mustafa, Farah
AU - Marquet, Roland
AU - Rizvi, Tahir Aziz
N1 - Funding Information:
This research was funded primarily by a grant from the United Arab Emirates University (UAEU) Program for Advanced Research-UPAR (UPAR-31M233) and in part by a grant from the College of Medicine and Health Sciences (31M280) to TAR. FNNP and AC were supported by UPAR-31M233 and UAE University Zayed Bin Sultan Center for Health Sciences (UCBR-31R123) grants, respectively. The authors would like to express their sincere thanks and appreciation to Dr. Mustafa T. Ardah and Dr. Syed Tariq for their help in gel filtration chromatography and electron microscopy, respectively.
Publisher Copyright:
© 2018, The Author(s).
PY - 2018/12/1
Y1 - 2018/12/1
N2 - MPMV precursor polypeptide Pr78Gag orchestrates assembly and packaging of genomic RNA (gRNA) into virus particles. Therefore, we have expressed recombinant full-length Pr78Gag either with or without His6-tag in bacterial as well as eukaryotic cultures and purified the recombinant protein from soluble fractions of the bacterial cultures. The recombinant Pr78Gag protein has the intrinsic ability to assemble in vitro to form virus like particles (VLPs). Consistent with this observation, the recombinant protein could form VLPs in both prokaryotes and eukaryotes. VLPs formed in eukaryotic cells by recombinant Pr78Gag with or without His6-tag can encapsidate MPMV transfer vector RNA, suggesting that the inclusion of the His6-tag to the full-length Pr78Gag did not interfere with its expression or biological function. This study demonstrates the expression and purification of a biologically active, recombinant Pr78Gag, which should pave the way to study RNA-protein interactions involved in the MPMV gRNA packaging process.
AB - MPMV precursor polypeptide Pr78Gag orchestrates assembly and packaging of genomic RNA (gRNA) into virus particles. Therefore, we have expressed recombinant full-length Pr78Gag either with or without His6-tag in bacterial as well as eukaryotic cultures and purified the recombinant protein from soluble fractions of the bacterial cultures. The recombinant Pr78Gag protein has the intrinsic ability to assemble in vitro to form virus like particles (VLPs). Consistent with this observation, the recombinant protein could form VLPs in both prokaryotes and eukaryotes. VLPs formed in eukaryotic cells by recombinant Pr78Gag with or without His6-tag can encapsidate MPMV transfer vector RNA, suggesting that the inclusion of the His6-tag to the full-length Pr78Gag did not interfere with its expression or biological function. This study demonstrates the expression and purification of a biologically active, recombinant Pr78Gag, which should pave the way to study RNA-protein interactions involved in the MPMV gRNA packaging process.
UR - http://www.scopus.com/inward/record.url?scp=85051214061&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85051214061&partnerID=8YFLogxK
U2 - 10.1038/s41598-018-30142-0
DO - 10.1038/s41598-018-30142-0
M3 - Article
C2 - 30087395
AN - SCOPUS:85051214061
SN - 2045-2322
VL - 8
JO - Scientific Reports
JF - Scientific Reports
IS - 1
M1 - 11793
ER -