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采用RNAiso-mate+改良CTAB法进行油桐花芽RNA的提取试验,用琼脂糖凝胶电泳检测RNA完整性,核酸蛋白质检测仪检测其纯度,研究提取油桐花芽高质量RNA的方法。结果表明:RNAiso-mate+改良CTAB方法提取的RNA质量好,条带清晰,亮度高,完整性及纯度较好,A260/A280比值为1.97、1.93,得率176.9、154.2μg/mL。在提取到的高质量RNA基础上,采用RT-PCR方法成功克隆了油桐SOC1基因片段,长度为611bp,含起始密码子,编码203个氨基酸;序列分析表明油桐SOC1基因与蓖麻、杨树、大豆的SOC1基因相似度分别为85%、84%、78%,进化树分析得出油桐SOC1基因与北美木兰SOC1基因亲缘关系较近。
RNAiso-mate + modified CTAB method was used to extract RNA from flower buds of Tung oil tree, RNA integrity was detected by agarose gel electrophoresis, the purity of RNA was detected by nucleic acid protein detector, and RNA extracted from Tung oilseed buds was studied. The results showed that RNAiso-mate + modified CTAB method had good quality, clear band, high brightness, good integrity and purity. The A260 / A280 ratio was 1.97,1.93, yield 176.9,154.2μg / mL. Based on the extracted high quality RNA, the SOC1 gene fragment of Tung tree was successfully cloned by RT-PCR with a length of 611bp and a start codon encoding a protein of 203 amino acids. The sequence analysis showed that the SOC1 gene of Castanea tung [...] The SOC1 gene similarity of poplar and soybean were 85%, 84% and 78% respectively. The phylogenetic tree analysis showed that the SOC1 gene of P. tung tree closely related to the SOC1 gene of Magnoliaceae.