与TMV-RNA装配起始位点互补的cDNA片段对TMV体外装配的抑制

合成了与TMV-RNA病毒装配起始位点互补的、长度为二十个核苷酸的DNA片段。该片段用~(32)P标记后,代替反义RNA(antisense RNA)与TMV-RNA进行硝基纤维素膜点杂交和溶液杂交。结果表明,该cDNA片段在两种条件下均能与TMV-RNA进行杂交。将溶液杂交的RNA-cDNA复合体经酒精沉淀,再与TMV衣壳蛋白的20S聚合体制剂进行体外装配,用测定310nm吸收光谱变化和电子显微镜观察的方法鉴定装配结果。实验证明,该cDNA片段与TMV-RNA杂交后抑制了装配起始位点的活力,从而使TMV病毒颗粒的装配不能完成。这一结果提示,TMV基因装配起始位点顺宁的cDNA和反义RNA能够在体外抑制TMV病毒颗粒的装配。

The cDNA of Assembly Origin Sequence in TMV Genome Inhibiting TMV Assembly in Vitro

A 20 nucleotides DNA fragment which is complementary with the assembly origin of TMV-RNA was synthesized. Instead of antisense RNA, the DNA fragment could hybridize with TMV-RNA in nitrocellulose dot hybridization and in mini-volume solution hybridization assays. Using RNA-cDNA complex and 20S aggregates of TMV coat protein, the TMV in vitro assembly reaction experiment, which was monitored by 310nm absorption spectrum and electron microscopy, showed that TMV in vitro assembly could not proceed in results of inhibition of the activity of assembly origin by the cDNA.