Sistem minigenom virus penyakit newcastle (NDV) AF2240 sebagai sistem pengekspresan gen sel mamalia

Translated title of the contribution: Minigenome system for newcastle disease virus (NDV) AF2240 as a gene expression system for mammalian cells

Shahrul Hisham Zainal Ariffin, Roslinda Shamsudin, Nurul Atikah Ahmad, Zulkiflie Zamrod

Research output: Contribution to journalArticle

Abstract

The minigenome system has been used as a model for studying transcription and replication for nonsegmented RNA viruses. The objective of this study was to develop a minigenome system for NDV local strain AF2240 and also to study the mechanism of replication and transcription of this virus. For this purpose, five recombinant plasmids; pMGNDV, pCITENP, pCITEP, pTriEX-T7 and pGEML were used. Transfection of all plasmids was carried out following large scale extraction and polyethylene glycol precipitation of plasmids. In vitro translational was performed by using pCITENP, pCITEP and pTriEX-T7 to ensure the constructs are functional. Western blot analysis showed that protein with approximate molecular weights of 100 kDa (T7), 53 kDa (NP), 53 and 55 kDa (P) was successfully detected. CAT protein was detected when plasmid encoding the NDV minigenome was cotransfected into BHK-21 cells with plasmid encoding nucleocapsid (NP), phosphoprotein (P) and large polymerase subunit (L) protein. Approximately, 55 pg CAT protein was successfully quantified by CAT ELISA. Western blot analysis also showed that the CAT protein with the size of 25 kDa was successfully obtained. In conclusion, this study showed that the system was functional and able to express foreign gene in mammalian cells BHK-21.

Original languageUndefined/Unknown
Pages (from-to)423-430
Number of pages8
JournalSains Malaysiana
Volume41
Issue number4
Publication statusPublished - Apr 2012

Fingerprint

Newcastle disease virus
Plasmids
Gene Expression
Nucleocapsid
Proteins
Western Blotting
RNA Viruses
Phosphoproteins
Protein Subunits
Virus Replication
Transfection
Molecular Weight
Enzyme-Linked Immunosorbent Assay
Genes

Keywords

  • Gene expression
  • Mammalian cell expression system
  • Minigenome system
  • Newcastle disease virus
  • Transfection

ASJC Scopus subject areas

  • General

Cite this

Sistem minigenom virus penyakit newcastle (NDV) AF2240 sebagai sistem pengekspresan gen sel mamalia. / Zainal Ariffin, Shahrul Hisham; Shamsudin, Roslinda; Ahmad, Nurul Atikah; Zamrod, Zulkiflie.

In: Sains Malaysiana, Vol. 41, No. 4, 04.2012, p. 423-430.

Research output: Contribution to journalArticle

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