水稻矮缩病毒基因组分析与部分基因片段的表达及功能研究  

Functional Genomic Analysis of Rice Dwarf Virus

作　　者：李毅[1] 徐洪[1] 程明非 郑宏红 冒志娟 肖锦[1] 张福建 明小天[1] 曲林[1] 刘一飞[1] 李玮[1] 赵晓岚 陈章良[1] 

机构地区：[1]北京大学生命科学学院

出　　处：《北京大学学报（自然科学版）》1998年第2期332-341,共10页Acta Scientiarum Naturalium Universitatis Pekinensis

基　　金：国家自然科学基金;欧盟(ERBCI-CT94-0088)资助

摘　　要：根据本实验室对水稻矮缩病毒（ＲｉｃｅＤｗａｒｆＶｉｒｕｓ，ＲＤＶ）中国福建分离株基因组全序列的测定，分析了其编码蛋白的功能，并将ＲＤＶＳ６，Ｓ９，Ｓ１０，Ｓ１１编码区ｃＤＮＡ分别克隆到表达载体ｐＴｒｃＨｉｓＡ，ｐＢＶ２２１，ｐＴｒｃＨｉｓＢ和ｐＢＶ２２１中，构建成表达质粒ｐＴＨ－Ｓ６，ｐＢＶＰ９，ｐＴＨ－Ｓ１０，ｐＢ－ＶＰ１１。分别用ＩＰＴＧ或温度诱导大肠杆菌ＤＨ５α，经ＳＤＳ－ＰＡＧＥ及相应抗体的Ｗｅｓｔｅｒｎｂｌｏｔｉｎｇ检测证明Ｓ６，Ｓ９，Ｓ１０，Ｓ１１编码蛋白在大肠杆菌中获得成功表达。另外，还将ＲＤＶ外层外壳蛋白基因Ｓ８、微核心蛋白基因Ｓ７及编码非结构蛋白基因Ｓ９通过基因枪导入到水稻植株中，抗病性正在研究之中。Rice dwarf virus full genome 12 gene segments were cloned and 25000 nucleotides were sequenced.The gene functions of each segment were analyzed according to amino acid sequences and experimental data.The coding regions of segments S6,S9,S10 and S11 encoding nonstractural proteins were cloned into expression vector pTricHisA,pBV221,pTric HisB,pBV221,respectively.The expressed products were analyzed by SDS PAGE and further confirmed by Western blotting.Using the E.coli expression products,the function of Pns11 was studied.The results showed that Pns11 is a ds /ss RNA and DNA binding protein and it may play very important roles in virus genome assortment or packaging.Gene segments S2,S7,S8 and S9 which encode virus most outer coat protein,minor core protein,outer coat protein and nonstructural protein were transferred into rice via biolistic bombardment and regenerated rice plants were obtained.The resistance to RDV infection of these transgenic rice plants are in testing.

关 键 词：基因组分析 原核表达 转基因水稻 水稻矮缩病毒 

分 类 号：S435.111[农业科学—农业昆虫与害虫防治]