![]() In low virulence-NDV has cleavage site that can only be split by enzymes that found only in the cell of digestive and respiratory tract. The amino acid sequence in the cleavage site is the main determinant of the viral virulence. On the surface of the virus particles, this protein is in the form of precursor Fo and can be activated if it split into F1 and F2 at the cleavage site. The F protein act to initiate the fusion of the viral envelopes and the host cell membranes. The HN protein involved in virus particle attachment to the sialic acid receptor on the surface of host cells. NDV infection process is depend on two major proteins, namely HN protein and F protein. The last is an asymptomatic enteric which is a subclinical enteric infection. Then NDV with a mild pathotype, namely lentogenic which is characterized by mild respiratory system disorders. Furthermore, the moderate pathotype, which is mesogenic, is characterized by respiratory and nervous system disorders with a low mortality rate. Velogenic neurotropic which is characterized by respiratory and nerve system disorders such as incoordination and torticollis which is characterized by high mortality rate. Based on the virulence and clinical symptoms in infected birds, NDV can be classified into five pathotypes, namely: viscerotropic velogenic which is a highly pathogenic NDV that is indicated by the haemorrhagic lesions in the intestinal mucosa with highly mortality rate. The viral particle of NDV is enveloped with the surface of viral particle covered by spike glycoproteins, namely Haemagglutinin-Neuraminidase (HN) protein and Fusion (F) protein. The genome of NDV is consist of negative single-strand RNA. Newcastle Disease is caused by NDV, which belongs to genus Avulavirus, family Paramyxoviridae and ordo mononegavirales. The outbreak of the disease can cause a high rate of mortality (up to 100%), which are very detrimental to the poultry farmers. Newcastle Disease is an infectious disease that infects many kinds of poultry and become a huge problem for the poultry industry around the world. This is an Open Access article distributed under the terms of the Creative Commons Attribution License 4.0, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. © The Authors, published by EDP Sciences, 2020 The visualisation of expressed recombinant F protein by SDS-PAGE and Westernblott showed the NDV recombinant F protein was a specific protein fragment with molecular weight of 25,6 kDa. coli contained plasmid pBT7-N-His (4.001 bp) inserted by recombinant F protein of NDV gene (642 bp). The expressed F protein of NDV then was visualized by SDS-PAGE and Westernblott to analyse the expression of NDV recombinant F protein. The recombinant F protein was in-vitro expressed by cell-free protein expression kit. This work started by detection of recombinant plasmid pBT7-N-His-Fusion NDV by DNA plasmid extraction followed by agarose gel electrophoresis. The aim of this study was to confirm whether clone C -2a of E.coli carrying a recombinant plasmid pBT7-N-His-Fusion NDV and to express a recombinant F protein of NDV in-vitro from expression plasmid by cell-free protein expression system. coli, then it was expressed by the Cell-free protein expression system. Fusion (F) protein encoding gene that has been inserted into pBT7-N-His expression p lasmid which isolated from clone C-2a of E. The previously study research reported that the local isolate of NDV from Kulon Progo, Indonesia has been isolated. Therefore, vaccination with new local NDV strain is needed to prevent the NDV infection in Indonesia. The antibody protection provided by the vaccine is not effective. These type of vaccine is 21%-23% genetically distinct with the virus that spread in the environment. NDV vaccine that used in Indonesia is a conventional life vaccine from LaSota and B1 strains. Vaccination is an effort to prevent the infection of NDV in poultry. Infection of NDV become a massive problem for poultry industry around the world especially in Indonesia. * Corresponding author: Disease Virus (NDV) is an infectious disease that infect many kinds of wild and domesticated birds. Ahmad Pandu Satria Wiratama and Aris Haryanto *ĭepartment of Biochemistry and Molecular Biology, Faculty of Veterinary Medicine, Universitas Gadjah Mada, Jl.Fauna 2 Karangmalang, Yogyakarta 55281, Indonesia
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