벼검은줄오갈병바이러스 외피단백질 유전자 단백질 발현과 항혈청 제작 (In Vitro Expression and Antibody Preparation of Rice black-streaked dwarf virus Coat Protein Gene)

서지정보

· 발행기관 : 한국식물병리학회
· 수록지 정보 : 식물병 연구 / 22권 / 1호 / 32 ~ 37페이지
· 저자명 : 이봉춘, 조상윤, 배주영, 김상민, 신동범, 김선림

초록

In this work, major outer capsid protein (P10) encoded by genome segment S10 of Rice blackstreakeddwarf virus (RBSDV) was expressed in Escherichia coli. Genomic dsRNA was extracted fromRBSDV-miryang isolate infected rice plants. Based on the sequence of S10 (RBSDV-miryang, GenBankJX994211), a pair of S10 specific primers were designed and used to amplify the fragment encodingthe N-part of P10. We amplified the partial gene (S10 1-834 nt) of RBSDV P10 (1-278 aa) by RT-PCR.
Amplified RBSDV S10 (1-834 nt) was cloned into the expression vector pET32a (+). RecombinantRBSDV S10 (1-834 nt) was expressed in E. coli BL21(DE3) and purified by nickel-nitrilotriacetic acid (Ni-NTA) affinity column. We successfully obtained P10 partial protein of RBSDV and the purified proteinwas used to immunize rabbits. The resulting polyclonal antiserum specifically recognized RBSDV frominfected plant in both Western blotting and enzyme-linked immunosorbent assay. In this study, weprovide purified RBSDV P10 (1-278 aa), which would be good material for the serological study ofRBSDV-miryang isolates.

영어초록

In this work, major outer capsid protein (P10) encoded by genome segment S10 of Rice blackstreakeddwarf virus (RBSDV) was expressed in Escherichia coli. Genomic dsRNA was extracted fromRBSDV-miryang isolate infected rice plants. Based on the sequence of S10 (RBSDV-miryang, GenBankJX994211), a pair of S10 specific primers were designed and used to amplify the fragment encodingthe N-part of P10. We amplified the partial gene (S10 1-834 nt) of RBSDV P10 (1-278 aa) by RT-PCR.
Amplified RBSDV S10 (1-834 nt) was cloned into the expression vector pET32a (+). RecombinantRBSDV S10 (1-834 nt) was expressed in E. coli BL21(DE3) and purified by nickel-nitrilotriacetic acid (Ni-NTA) affinity column. We successfully obtained P10 partial protein of RBSDV and the purified proteinwas used to immunize rabbits. The resulting polyclonal antiserum specifically recognized RBSDV frominfected plant in both Western blotting and enzyme-linked immunosorbent assay. In this study, weprovide purified RBSDV P10 (1-278 aa), which would be good material for the serological study ofRBSDV-miryang isolates.

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