Cloning of hexon gene of egg drop syndrome virus and construction of its eukaryotic expression vector

Wu ZhiMing; Zhang ChunJie; Li YinJu; Cheng XiangChao

Journal of Henan Agricultural University 40(1): 7-10

2006


Accession: 015292366

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Abstract
Hexon gene of egg drop syndrome virus (EDSV) was amplified from EDSV by a touchdown polymerase chain reaction (TD-PCR). It was cloned into the vector pMD18-T and then transformed into competent cells of Escherichia coli str. JM109. Furthermore, the cloned gene was inserted into eukaryotic expression vector pcDNA 3 to obtain recombinant vector pcDNA 3-hexon, and transformed into E. coli str. JM109. The pcDNA 3-hexon was subjected to the enzyme digestion identification and sequencing. The full length of hexon gene was 2.733 kb, encoding 910 amino acids. Enzyme digestion identification and sequencing showed the inserted sequence in the expression vector pcDNA 3-hexon was identical to amplified hexon gene sequence, with a correct open reading frame. It is concluded the eukaryotic expression vector pcDNA 3-hexon was successfully constructed.