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| Preliminary Study of a New Transient Oviduct Bioreactor Mediated by Recombinant Avian Adeno-associated Virus |
| WANG An-ping1,SUN Huai-chang2 |
1.Jiangsu Animal Husbandry and Veterinary College,Taizhou 225300,China
2.College of Veterinary Medicine,Yangzhou University,Yangzhou 225009,China |
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Abstract To research a new transient oviduct bioreactor expressing recombinant human tissue kallikrein,hKLK1 cDNA-expression cassette was subcloned into avian adeno-associated virus transfer vector pAITR,and transfected into AAV-293 cells with AAAV helper vector pcDNA-ARC and adenovirus helper vector pHelper using calcium phosphate precipitation method.The recombinant AAAV-KLK1 were injected into laying hens via wing vein at 1×1010 virus particles per hen.Oviduct-specific expression of hKLK1 was demonstrated by reverse transcription-PCR; the enzymatic activity results showed that the kallikrein expression could be detected at the second day after injection,reaching the highest level of 107.3U/ml at the third week,and lasted for 6 weeks after injection; Oral administration of hKLK-contained egg white had significant hypotensive effect on spontaneous hypertension rats，which lasted for 5 days. These data suggest that the transient oviduct reactor mediated by recombinant avian adeno-associated virus can be used for high level and long-lasting hKLK1 expression,and the production of other recombinant proteins.
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Received: 18 May 2009
Published: 21 December 2009
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