論文

2019年6月1日

Expression of the thermostable Moloney murine leukemia virus reverse transcriptase by silkworm-baculovirus expression system

Journal of Asia-Pacific Entomology
  • Takumi Yano
  • ,
  • Jae Man Lee
  • ,
  • Jian Xu
  • ,
  • Yoshiki Morifuji
  • ,
  • Akitsu Masuda
  • ,
  • Masato Hino
  • ,
  • Daisuke Morokuma
  • ,
  • Ryosuke Fujita
  • ,
  • Masateru Takahashi
  • ,
  • Takahiro Kusakabe
  • ,
  • Hiroaki Mon

22
開始ページ
453
終了ページ
457
DOI
10.1016/j.aspen.2019.02.008

© 2019 Korean Society of Applied Entomology Reverse transcriptase from Moloney murine leukemia virus (MMLVRT) is an RNA dependent DNA polymerase, which has been used as a fundamental tool for molecular biology and biotechnology. The secondary structures formed in the RNA templates decrease the accessibility of the reverse transcriptase to the RNA templates; it is important to unfold the RNA secondary structure by increasing the reaction temperature to perform the successful transcription. In this study, we applied silkworm baculovirus expression vector system (silkworm-BEVS) to mass-produce and purify the recombinant MMLVRTs with the N- or C- terminal tandem tags. We confirmed that both of the recombinant MMLVRT enzymes have intact DNA polymerase activity. It is notable that C- terminal tagged MMLVRT outperformed MMLVRT obtained from the E. coli expression system in terms of thermostability and sensitivity to low quantities of RNA template. Taken together, these results demonstrate that silkworm-BEVS is a promising alternative strategy to produce the functional and thermostable reverse transcriptase.

リンク情報
DOI
https://doi.org/10.1016/j.aspen.2019.02.008
Scopus
https://www.scopus.com/record/display.uri?eid=2-s2.0-85062422843&origin=inward
URL
https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85062422843&origin=inward