Progress 01/15/04 to 01/14/08
Outputs
OUTPUTS: We have constructed a cDNA clone encoding the entire 14,150-nucleotide genome of AMPV/CO by assembling five cDNA fragments into a transcription plasmid, pBR 322.
PARTICIPANTS: Samal, S.K. - Planning of the project. Govindarajan, D. - Performing required experiments, etc.
TARGET AUDIENCES: Poultry industry
PROJECT MODIFICATIONS: None
Impacts
Transfection of this plasmid, along with expression plasmids encoding the N, P, M2-1 and L proteins of AMPV/CO, into cells stably expressing T7 RNA polymerase resulted in the recovery of infectious AMPV/CO. Characterization of the recombinant AMPV/CO showed that its growth properties in tissue culture were similar to those of the parental virus. These studies completed the proposed objectives from our previous USDA/NRI project. Furthermore, we have evaluated the potential of AMPV/CO to serve as a viral vector by generating another recombinant virus, rAMPV/CO-GFP, that expressed GFP as a foreign protein.
Publications
- Govindarajan, D. and Samal, S.K. (2004) Sequence analysis of the large polymerase (L) gene of the US strain of avian pneumovirus indicates a close resemblance to that of human metapneumovirus. Virus Res. 105(1): 59-66
- Govindarajan, D., Yunus, A.S. and Samal, S.K. (2004) Complete sequence of the G glycoprotein gene of avina metapneumovirus subgroup C and identification of a divergent domain in the predicted protein. J. Gen. Virol. 85:3671-3675
- Govindarajan, D. and Samal, S.K. (2005) Analysis of the complete genome sequence of avian metapneumovirus subgroup C indicates that it possesses the longest genome among metapneumoviruses. Virus Genes 30:329-331
- Govindarajan, D., Buchholz, U.J. and Samal, S.K. (2006) Recovery of an avian metapneumovirus subgroup C from cDNA: Cross-recognition of avian and human metapneumovirus support proteins, J. Virol. 80(12): 5790-5797
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Progress 01/01/05 to 12/31/05
Outputs
A cDNA clone encoding the entire 14,150nt genome of strain AMPV/CO was generated by assembling five cDNA fragments between the T7 RNA polymerase promoter and the autocatalytic hepatitis delta virus ribosyme of a transcription plasmid pBR 322. Transfection of this plasmid, along with the expression plasmids encoding the N,P,M2-1 and L proteins of AMPV/CO, into cells stably expressing T7 RNA polymerase resulted in the recovery of infectious AMPV/CO. Characterization of the recombinant AMPV/CO showed that its growth properties in tissue culture were similar to that of the parental virus. The potential of AMPV/CO to serve as a viral vector was also assessed by generating another recombinant virus, rAMPV/CO-GFP, that expressed the enhanced green fluorescent protein (GFP) as a foreign protein. Interestingly, GFP-expressing AMPV and GFP-expressing human metapneumovirus (HMPV) could be recovered using the support plasmids of either virus, denoting that the genome promoters
are conserved between the two metapneumoviruses and can be cross-recognized by the polymerase complex proteins of either virus. These results indicate a close functional relationaship between AMPV/CO and HMPV.
Impacts
This newly-developed reverse genetics system will be very helpful in studying the basic molecular biology of avian metapneumoviruses and in developing attenuated live recombinant vaccines to control this emerging poultry pathogen. Furthermore, this system can also be used to develop vectored vaccines for other avian pathogens. Therefore, these studies will be greatly beneficial to the poultry industry.
Publications
- Govindarajan, D. and Samal, S.K. 2004. Sequence analysis of the large polymerase (L) protein of the US strain of avian metapneumovirus indicates a close resemblance to that of the human metapneumovirus. Virus Res. 105:59-66.
- Govindarajan, D., Yunus, A.S. and Samal, S.K. 2004. Complete sequence of the G glycoprotein gene of avian metapneumovirus subgroup C and identification of a divergent domain in the predicted protein. J Gen Virol. 85:3671-3675.
- Govindarajan, D. and Samal, S.K. 2005. Analysis of the complete genome sequence of avian metapneumovirus subgroup C indicates that it possesses the longest genome among metapneumoviruses. Virus Genes 30:331-333.
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