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ACTA HORTICULTURAE SINICA ›› 2017, Vol. 44 ›› Issue (10): 2001-2007.doi: 10.16420/j.issn.0513-353x.2016-0826

• Research Notes • Previous Articles     Next Articles

Prokaryotic Expression of Basella rugose mosaic virus CP Gene and Preparation of Its Polyclonal Antiserum

LI Xiaoqin1,ZHANG Pengyuan1,2,REN Longhui1,3,DAI Jinran1,WANG Jianguang1,*,and CHEN Suiyun1,*   

  1. (1School of Life Sciences,Yunnan University,Kunming 650091,China;2School of Life Science,Southwest Forestry University,Kunming 650224,China;3Yunnan Oriental Tobacco Co.,Ltd.,Baoshan,Yunnan 678000,China)
  • Online:2017-10-25 Published:2017-10-25

Abstract: The full length cp gene of Basella rugose mosaic virus(BaRMV)was obtained by RT-PCR in the infected Mirabilis jalapa L. leaves and the prokaryotic experssion vector of BaRMV cp gene was successfully constructed also. The pET30a-BaRMV-cp vector was transferred into Rosetta(DE3)PlysS. SDS-PAGE results indicated that the His-CP fusion protein was about 40.0 kD in size,which was in accord with the prediction. Target proteins were isolated by cutting the gel slices that contained the 40.0 kD bands which were stained by 0.25 mol ? L-1 KCl. Purified fusion protein was used to immunize New Zealand white rabbits to produce the antiserum. The polyclonal antiserum was detected with indirect enzyme-linked immunosorbent assay(ID-ELISA),the titer of the antiserum was1︰16 000,Western-blotting analysis showed that the antiserum has very strong specificity. Detected the field susceptible M. jalapa L. and the laboratory inoculated and infectious Nicotiana benthamiana L. by the antiserum,the test result also showed that the prepared antiserum has good specificity and can be used for large-scale sample testing.

Key words: Basella rugose mosaic virus(BaRMV), Mirabilis jalapa, coat protein gene, polyclonal antiserum, ID-ELISA, Western blotting

CLC Number: