LUO Min,SUN Ying,CHEN Xian,et al.Study on Improvement of Methods for RNA Isolation From Flower Bud of Vernicia fordii and Clone and Analysis of VfSOC1?Gene[J].Northern Horticulture,2014,38(14):82-85.
油桐花芽RNA提取方法的改良及油桐SOC1基因
- Title:
- Study on Improvement of Methods for RNA Isolation From Flower Bud of Vernicia fordii and Clone and Analysis of VfSOC1?Gene
- 文章编号:
- 1001-0009（2014）14-0082-04
- Keywords:
- Vernicia fordii; flower bud; RNA extraction; SOC1 gene
- 分类号:
- S 794.3
- 文献标志码:
- A
- 摘要:
- 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、154.2 μg/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611 bp，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。
- Abstract:
- Using RNAiso-mate+modified CTAB method for RNA extraction of Vernicia fordii flower bud and detect RNA and protein by agarose gel electrophoresis and nucleic acid protein detector respectively,the method of RNA extraction of Vernicia fordii flower bud were studied.The results showed that the RNA of high quality,clear strip,high brightness,good integrity and purity was extracted by the method of RNAiso-mate+modified CTAB.The ratio of A260 and A280 was 1.97 and 1.93 respectively,yield was 176.9 and 154.2 μg/mL respectively.Based on the extracted high quality RNA,the VfSOC1?gene fragment was successfully cloned by reverse transcription polymerase chain reaction (RT-PCR).The length of VfSOC1 gene fragment was 611 bp and encode 203 amino acids.Compared with SOC1genes of Ricinus communis,Popμlus trichocarpa,Glycine max, VfSOC1 share 85%,84% and 78% identity respectively.The phylogenetic analysis showed that the genetic relationship of VfSOC1 and MvSOC1 was closer.
参考文献/References:
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[2]曾凡锁，南楠，詹亚光，富含多糖和次生代谢产物的白桦成熟叶中总RNA的提取[J]．植物生理学通讯，2007，43(5)：913-916．
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备注/Memo
第一作者简介：罗敏（1987-），女，硕士研究生，研究方向为经济林栽培育种。E-mail：290156927@qq.com.