香蕉束顶病毒复制酶基因克隆及转基因表达

The Cloning of Replicase Gene of Banana Bunchy Top Virus and the Gene Expression

摘要:

以广州市郊获得的香蕉束顶病毒(BBTV)的DNA为模板,进行PCR扩增得到香蕉束顶病毒复制酶基因的1.1 kb DNA。所获得的DNA序列与澳大利亚的BBTV序列的同源性达90%,这部分序列编码香蕉束顶病毒复制酶基因的羧基端。将改造的BBTV复制酶基因克隆到pBI 121的CaMV35S和NOS终止序列之间,构建表达载体,并采用基因枪轰击香蕉试管苗生长点组织的方法,经PCR检测和Westernblot分析,获得4株具有BBTV复制酶基因整合表达的T₀代转基因香蕉。转基因植株的抗病性正在检测之中。

Abstract:

The DNA was isolated from banana grown in Guangzhou, which was infected with bunchy top virus. 1.1 kb DNA of replicase of banana bunchy top virus (BBTV) was obtained by PCR using this virus DNA astemplates for amplification. The derived DNA of the BBTV replicase was encoded by the C-Terminal of BBTV. It was homologous to 90% with the Australian BBTV. The reformed BBTV replicase was cloned to pBI 121 in the position between CaMV 35S promoter and NOS termination sequence, and a plant expressed carrier was established. Four transgenic platelets with BBTV replicase gene in T₀ generation were detected by PCR and Western blot analysis. The ability of these transgenic banana against bunchy top virus is being investigated.