[Objective] Aimed to construct RNAi vector resistant to cucumber mosaic virus and transferred this vector into tobacco. [Method] RT-PCR method was used to amplify cucumber mosaic virus NS04 and process RNA2 gene sequen of tomato isolates. The analysis results of phylogenetic tree demonstrated that the sequence in RNA2 encoded CMV-2a had 98.0% and 96.5% homology with nucleotide and amino acid of DQ412731 isolate of Zhejiang,China. The replicase fragment in CMV RAN2 gene was taken as target sequence to construct pBi35SCR2 eukaryotic expression vector,then the expression vector was identified. Through agrobacterium-mediated method,the expression vector was transferred into tabacco and PCR method was used to check the transfer. The PCR results demonstrated that the experiment had successfully construct eukaryotic expression vector of pBi35SCR2 and the expression vector was successfully transferred into tabacco. [Conclusion] The obtained transgenic tobacco could be used as challenge test material in following experiment and provided foundation for studying processing tomato resist cucumber mosaic virus.
【目的】海岛棉成花素类似基因的克隆及生物信息学分析。【方法】利用RT-PCR技术,从海岛棉中克隆成花素FLOWERING LOCUS T(FT)的同源基因,并进行生物信息学分析。【结果】从新海14号花后15 d纤维中,克隆到一个棉花成花素类似基因FLOWERING LOCUS T-LIKE 1,命名为GbFTL1。该基因的开放阅读框为525 bp,编码174个氨基酸。蛋白比对表明GbFTL1和AtFT的相似性为79.3%,和陆地棉GhFTL1的相似性为99.4%,GbFTL1第37位氨基酸为Asn(N),而GhFTL1第37位氨基酸为Ser(S)。GbFTL1含有FT蛋白亚家族两个关键的氨基酸残基及14个保守氨基酸,系统进化树分析表明GbFTL1属于FT亚家族成员。【结论】GbFTL1基因可能为海岛棉中促进开花的基因之一。
