ABSTRACT
Aim: To investigate molecular and evolutionary characteristics of genes of fowlpox virus (FWPV) isolates from chickens in Tanzania. Study Design: Experimental. Place and Duration of Study: Faculty of Veterinary Medicine, Sokoine University of Agriculture, Morogoro, Tanzania; between November 2011 and October 2013. Methodology: Samples of cutaneous nodular lesions were collected from featherless parts of chickens (n = 154) suspected to have fowl pox in 14 regions of Tanzania followed by virus isolation, DNA extraction, polymerase chain reaction (PCR) amplification of the P4b gene, gel electrophoresis of PCR products, purification of PCR products, sequencing of purified PCR products and finally analysis of sequence data using standard procedures. Results: The disease was confirmed in 12 regions, out of 154 investigated samples 66 (42.86%) were found to contain FWPV, indicating that the 66 chickens from which the samples were collected had fowl pox as a result FWPV infection. Sequence analysis revealed that the Tanzanian FWPV isolates were 99.65 – 100% identical to each other and 99 – 100% identical to several published sequences of FWPV isolates from various countries in different continents of the world, including Europe and Asia. Phylogenetic analysis revealed that all Tanzanian isolates belong to clade A, subclade A1. Conclusion: Based on the findings of this study it is concluded that currently fowl pox is prevalent in several regions of Tanzania, caused by FWPVs which are genetically and phylogenetically closely related. However, these findings do not rule out the possibility of existence of genetic divergence among FWPVs currently prevalent in Tanzania. In order to rule out or detect genetic divergence (if any) among FWPVs currently prevalent in the country, other studies aimed at investigating molecular and evolutionary characteristics of genes in other genomic regions are highly recommended.
ABSTRACT
Objective:To express Gag gp120 fusion protein in the recombinant fowlpox virus.Methods:The Gag gp120 gene of HIV 1 was inserted downstream of the combined promoter in pUTAL,an expressing plasmid was constructed.By transfecting the plasmid into CEF cells pre infected with FPV 282E 4 strain,a recombinant fowlpox virus vUTALG was obtained by selecting in the presence of BUdR and subsequent blue plaque screening.Results:Western blot showed that the recombinant virus expressed the Gag gp120 fusion protein in the infected CEF cells.The virus like particles formed by Gag protein were observed under electron microscope.Mice were immunized with the recombinant virus.The results showed the recombinant virus could induce HIV 1 specific antibody response.Conclusion:The recombinant fowlpox virus could express the Gag gp120 fusion protein.
ABSTRACT
Objective To provide experimental evidences for developing a safe and effective re- combinant fowlpox virus which can prevent the infection of HIV-2.Methods A fowlpox virus(FPV) transferring vector was constructed by inserting HIV-2 gag gene to the downstream of a synthetic complex promoter ATI-p7.5?20 of vector pUTA2.Transfection was then carried out,and recombi- nant FPV(rFPV)was screened by 5'-bromo-deoxyuridine(BrdU),genome PCR and western blot detection.Balb/c mice were immunized with rFPV by muscular injection.Anti-HIV-2 antibody, CD4~+ and CD8~+ T-cell count and specific target-killing activity of spleen CTL in immunized mice were analyzed by ELISA,FACS and LDH release assay,respectively.Results A transferring vector pA- gag was constructed and confirmed by amplifying a fragment of 766 bp from the rFPV genome.Mean- while,HIV-2 multi-antibody-specific protein blot(55 000)was detected from the recombinant virus and the HIV-2 specific antibody was detected from the immunized Balb/c mice.HIV-2 specific target- killing activity of spleen CTL was observed in immunized mice.Conclusion A recombinant fowlpox virus expressing HIV-2 structural protein Gag has been obtained,and it can stimulate HIV-2-specific eelluar and humoral immune reactions in mice.