A pair of primers was designed based on the vp28 gene and PCR was performed to amplify the gene from WSSV DNA. Inserting the DNA fragment to the yeast-E.coli shuffle vector pPICZ and the recombinant plasmid (pPICZVP28...A pair of primers was designed based on the vp28 gene and PCR was performed to amplify the gene from WSSV DNA. Inserting the DNA fragment to the yeast-E.coli shuffle vector pPICZ and the recombinant plasmid (pPICZVP28) that contains the target DNA fragment was obtained in the E.coli strain.The pPICZVP28 was introduced into Pichia pastoris strain X33 by electroporation. The transformant strain X33-1 was grown in BMGY media in fled flask at 28℃. Induced by methanol for 72h, the samples of cell pellets and supernatant were collected by centrifugation and analyzed by SDS-PAGE and Western-blot, which confirmed that the strain X33-1 can express the WSSV’s envelope protein vp28.展开更多
文摘A pair of primers was designed based on the vp28 gene and PCR was performed to amplify the gene from WSSV DNA. Inserting the DNA fragment to the yeast-E.coli shuffle vector pPICZ and the recombinant plasmid (pPICZVP28) that contains the target DNA fragment was obtained in the E.coli strain.The pPICZVP28 was introduced into Pichia pastoris strain X33 by electroporation. The transformant strain X33-1 was grown in BMGY media in fled flask at 28℃. Induced by methanol for 72h, the samples of cell pellets and supernatant were collected by centrifugation and analyzed by SDS-PAGE and Western-blot, which confirmed that the strain X33-1 can express the WSSV’s envelope protein vp28.
