The genome segment S7 of Rice blackstreaked dwarf virus (RBSDV) contains two nonoverlapping major open reading frames (ORFs), which encoded two proteins, p7a and p7b,respectively. Based on the sequence of S7 (EU111804), two pairs of S7specific primers were designed and respectively used to amplify the fragments encoding the p7a and p7b proteins. The expected products were subcloned into the expression vector pET32a（+） or pSBET and expressed at high level in E. coli BL21(DE3)pLysS or BL21(DE3)pLysE, respectively. An antiserum specifically against p7a or p7b proteins was prepared using the purified expression proteins to immunize mice. The results of Western blot revealed that both p7a and p7b proteins were expressed in infected plants, and p7a protein was more abundant and easily detected than p7b protein. However, both have not been detected in the viral particles, indicating that both proteins encoded by RBSDV S7 were nonstructural proteins.
LU Mingfang1,2, YANG Jian2, ZHANG Hengmu2, CHEN Jianping2,*
. Detection of Two Nonstructural Proteins Encoded by Genome Segment S7 of Rice BlackStreaked Dwarf Virus in Infected Rice Plants[J]. Chinese Journal OF Rice Science, 2012
, 26(1)
: 9
-15
.
DOI: 10.3969/j.issn.10017216.2012.01.003
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