Kim Hui-Bae, Kim Do-Yeong, Cho Tae-Ju
Department of Biochemistry, Chungbuk National University, Cheongju 361-763, Korea.
BMB Rep. 2014 Jun;47(6):330-5. doi: 10.5483/bmbrep.2014.47.6.187.
Turnip yellow mosaic virus (TYMV) is a spherical plant virus that has a single 6.3 kb positive strand RNA as a genome. In this study, RNA1 sequence of Flock house virus (FHV) was inserted into the TYMV genome to test whether TYMV can accommodate and express another viral entity. In the resulting construct, designated TY-FHV, the FHV RNA1 sequence was expressed as a TYMV subgenomic RNA. Northern analysis of the Nicotiana benthamiana leaves agroinfiltrated with the TY-FHV showed that both genomic and subgenomic FHV RNAs were abundantly produced. This indicates that the FHV RNA1 sequence was correctly expressed and translated to produce a functional FHV replicase. Although these FHV RNAs were not encapsidated, the FHV RNA having a TYMV CP sequence at the 3'-end was efficiently encapsidated. When an eGFP gene was inserted into the B2 ORF of the FHV sequence, a fusion protein of B2-eGFP was produced as expected.
芜菁黄花叶病毒（TYMV）是一种球形植物病毒，其基因组为一条6.3 kb的单链正义RNA。在本研究中，将禽传染性支气管炎病毒（FHV）的RNA1序列插入TYMV基因组，以测试TYMV是否能够容纳并表达另一种病毒实体。在所得的构建体（命名为TY-FHV）中，FHV RNA1序列作为TYMV亚基因组RNA进行表达。对用TY-FHV进行农杆菌浸润的本氏烟草叶片进行Northern分析表明，基因组FHV RNA和亚基因组FHV RNA均大量产生。这表明FHV RNA1序列被正确表达并翻译以产生功能性FHV复制酶。尽管这些FHV RNA未被衣壳化，但在3'-末端具有TYMV CP序列的FHV RNA被有效地衣壳化。当将eGFP基因插入FHV序列的B2 ORF中时，预期产生了B2-eGFP融合蛋白。
