Cloning and expression of the phosphoprotein gene of Newcastle disease virus in Escherichia coli.
The phosphoprotein (P) gene of a heat stable Newcastle disease virus (NDV) was cloned, sequenced and expressed in Escherichia coli. SDS-PAGE analysis of the recombinant P protein (395 amino acids) and a C-terminal extension derivative (424 amino acids), gave rise to two distinct protein bands with molecular masses of approximately 53-55 and 56-58 kDa, respectively, which are approximately 26-30% heavier than those calculated from the deduced amino acid sequences. The differences in molecular mass on SDS-PAGE are thought to be attributed to the acidic nature of the P protein (pI=6.27) and also the different degrees of phosphorylation in the prokaryotic cell. Amino acid sequence comparison of the P protein among the published NDV strains showed that they were highly conserved particularly at the putative phosphorylation sites.
Kho
Chiew Ling
CL
Department of Biochemistry and Microbiology, Faculty of Science and Environmental Studies, Universiti Putra Malaysia, 43400 UPM, Serdang, Selangor, Malaysia.
Tan
Wen Siang
WS
Yusoff
Khatijah
K
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004279:DNA, Viral / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005814:Genes, Viral; D007526:Isoelectric Point; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009522:Newcastle disease virus / Q000235:genetics*; D010750:Phosphoproteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D010766:Phosphorylation; D010957:Plasmids / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014764:Viral Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*
D004279:DNA, Viral; C055691:P phosphoprotein, Newcastle disease virus; D010750:Phosphoproteins; D011994:Recombinant Proteins; D014764:Viral Proteins