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| Rapid construction of multiple target RNAi expression vector from synthesized fragment |
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| DOI：10.16768/j.issn.1004-874X.2016.07.013 |
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| Abstract: |
| Papaya production is seriously limited by Papaya ringspot virus（ PRSV） worldwide and Papaya leaf-distortion mosaic virus（ PLDMV） in Eastern Asia. To develop transgenic papaya lines with robust resistance to both of PRSV and PLDMV viruses， we took advantage of a sythesized fragment which included several conserved segments of Nib and Hc-Pro gene of PRSV and PLDMV to constructe pPTN-LS RNAi expression vector. The pPTNLS RNAi expression vector was constructed by two fragments with different length and inverted repeat（ IR） deriving from the sythesized fragment. The IR part served as stem while the extending part of the longer fragment served as loop
of the hairpin structure of pPTN-LS respectively. Since it didn’t need to insert additional intron，the method of RNAi
expression vector construction mentioned above was fast and the result was stable as well. The transgenic Papaya engineering with pPTN-LS should be robust resistant to PRSV and PLDMV at the same time，because the sythesized fragment was derived from the highly conserved regions of Nib and Hc-Pro gene of PRSV and PLDMV. The study maypave the way to develop transgenic papaya lines with robust resistance to both of PRSV and PLDMV viruses. |
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