Cloning and expression analysis of multiple proteins encoding P gene of Newcastle disease virus.
Viral gene oncotherapy is emerging as a biotherapeutic cancer treatment modality based on targeted killing of cancer cells by viral genes. Newcastle disease virus (NDV) has the property to cause selective oncolysis of tumor cells sparing normal cells. NDV has a single stranded negative sense RNA genome, which is 15,186 nucleotide long and consists of six genes, which codes for eight proteins. NDV like other paramyxoviruses has the ability to generate multiple proteins from the P gene. P protein is encoded by an unedited transcript of the P gene, whereas the V and W protein are the results of RNA editing event in which one and two G residues are inserted at a conserved editing site within the P gene mRNA resulting in V and W transcripts, respectively. Although NDV is known to cause oncolysis by triggering apoptosis, the role of different viral proteins in selective oncolysis is still unclear. P gene edited products are known for its anti-apoptotic property in homologous host. In the present study, NDV P gene and its RNA edited products were amplified, cloned, sequenced and in vitro expression was done in HeLa cells. Further constructs were assayed for their apoptosis inducing ability in HeLa cells. Preliminary study suggested that P, V and W proteins are not apoptotic to HeLa cells.
Molecular Biology Laboratory, Department of Veterinary Biotechnology, Indian Veterinary Research Institute, Izatnagar 243 122, India.
Rajiv
R
Kumar
Ashok K
AK
Tiwari
Uttara
U
Chaturvedi
G Ravi
GR
Kumar
A P
AP
Sahoo
Sudesh
S
Kumar
Sangeeta
S
Tiwari
D000595:Amino Acid Sequence; D000818:Animals; D017304:Annexin A5 / Q000378:metabolism; D001483:Base Sequence; D002645:Chickens; D003001:Cloning, Molecular; D015967:Gene Expression Regulation, Viral; D005814:Genes, Viral / Q000235:genetics*; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D009522:Newcastle disease virus / Q000235:genetics*; D016366:Open Reading Frames / Q000235:genetics; D010750:Phosphoproteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D015203:Reproducibility of Results; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*
D017304:Annexin A5; C055691:P phosphoprotein, Newcastle disease virus; D010750:Phosphoproteins; D014764:Viral Proteins