甘薯和巴西牵牛18S rRNA基因的克隆和序列分析

Cloning and Sequence Analysis of 18S rRNA Gene in Ipomoea batatas and Ipomoea setosa

【研究目的】为甘薯和侵染甘薯病毒的基因表达研究提供内参基因序列信息。【方法】分别以‘商薯19’、‘北京553’2个甘薯品种和巴西牵牛的基因组DNA为模板,利用PCR方法克隆甘薯和巴西牵牛18S rRNA基因序列。【结果】测序结果表明,获得的供试2甘薯品种和巴西牵牛的18S rRNA基因序列长度均为1630bp;序列比对结果表明,甘薯和巴西牵牛与裂叶牵牛、烟草等双子叶植物的18S rRNA基因相应序列的一致性均达98%以上,与单子叶植物Lilium superbum的18S rRNA基因序列也有较高的一致性。【结论】从2甘薯品种和巴西牵牛的基因组中克隆出了18S rRNA基因部分序列,研究结果不仅为利用18S rRNA基因作为内参基因分析甘薯和侵染甘薯病毒基因的表达研究提供了序列依据,而且可为甘薯和巴西牵牛的分子系统学研究提供序列参考。

[Objective] to supply sequence information of internal control gene for analyzing gene expression of I.batatas and viruses infectingI.batatas .[Method] Sequences of 18S rRNA gene were cloned using PCR method from genomic DNA ofI.batatas cultivar‘Shangshu19’,‘Beijing553’andI.setosa,respectively.[Result] The sequencing of the DNA fragments all generated a total of 1630 bp nucleotide sequence.The obtained 18S rRNA gene sequences ofI.batatas andI.setosa shared more than 98% identity withI.hederacea and Nicotiana tabacum among dicotyledons,and shared high identity with Lilium superbum among monocotyledons.[Conclusion] Partial sequences of 18S rRNA gene were cloned from genomic DNA of I.batatas cultivar‘Shangshu19’,‘Beijing553’andI.setosa,which provided sequence information not only for analyzing gene expression ofI.batatas and viruses infectingI.batatas using 18S rRNA gene as internal control,but for molecular systematic research ofI.batatas andI.setosa.