中国农业科学院农业信息研究所 中国农学会
Agricultural Information Institute of CAAS China Association of Agricultural Science Societies
En
Ch
中国农业科学院农业信息研究所 中国农学会 主办

AI检索

订阅

返回顶部
当前位置: 首页 > 文章 > 枇杷果实总RNA的提取及八氢番茄红素合成酶基因(PSY)片段的克隆 果树学报 2012 (4) 59-64
Position: Home > Articles > Extraction of total RNA from loquat fruit and cloning of phytoene synthase gene fragment Journal of Fruit Science 2012 (4) 59-64

枇杷果实总RNA的提取及八氢番茄红素合成酶基因(PSY)片段的克隆

作  者：
张玲;林顺权;杨向晖
单  位：
华南农业大学园艺学院
关键词：
枇杷果实;总RNA提取;PSY基因
摘  要：
【目的】为从枇杷果实中提取高质量的RNA,【方法】以枇杷果实为材料,对6种总RNA提取方法进行分析和比较。【结果】结果表明,在裂解前先用70%丙酮和80%乙醇分别对枇杷果皮和果肉进行预处理,再结合CTAB-LiCl法,得到的总RNA质量较好,OD260/OD280在1.8～2.2,OD260/OD230超过2.0,得率分别为幼果果皮56.78μg.g-1,幼果果肉40.62μg.g-1,成熟果果皮40.86μg.g-1,成熟果果肉9.24μg.g-1;并根据其他植物的八氢番茄红素合成酶基因(PSY)保守区设计引物,用以上方法提取的RNA为模板进行RT-PCR,得到453 bp的基因片段,其推导的氨基酸序列与其他植物的同源性高达95%,推测该片段为PSY基因。【结论】先用70%丙酮和80%乙醇分别对枇杷果皮和果肉进行预处理,再结合CTAB-LiCl法,得到的RNA质量好,可用于后续的分子生物学研究。
译  名：
Extraction of total RNA from loquat fruit and cloning of phytoene synthase gene fragment
作  者：
ZHANG Ling,LIN Shun-quan,YANG Xiang-hui(College of Horticulture,South China Agricultural University,Guangzhou,Guandong 510642,China)
关键词：
Loquat fruit;Total RNA extraction;PSY gene
摘  要：
【Objective】 The objective of the study is to isolate high quality of RNA from loquat fruit.【Method】 Six RNA extraction methods were analyzed and compared.【Result】 The results showed that the total RNA with desirable yield and quality can be extracted with CTAB-LiCl protocol with preprocessing peel and pulp of fruit by 70% acetone and 80% alcohol before lysis respectively.The OD260/OD280 were among 1.8 and 2.2,OD260/OD230 were higher than 2.0,and the yields of peel of young fruit,pulp of young fruit,peel of ripe fruit,pulp of ripe fruit were 56.78 μg·g-1,40.62 μg·g-1,40.86 μg·g-1 and 9.24 μg·g-1 respectively.In addition,according to conserving sequence of phytoene synthase(PSY) gene from several other plants,the primers were designed for the reverse transcription-polymerase chain reaction(RT-PCR),and the 453 bp fragment of loquat was obtained.【Conclusion】 The studies suggested that the total RNA with desirable yield and quality can be extracted with CTAB-LiCl protocol with preprocessing peel and pulp of fruit by 70% acetone and 80% alcohol respectively,and the extracted RNA could be used in further molecular research.

相似文章

导出引用
计量
文章访问数: 7
HTML全文浏览量: 0
PDF下载量: 0

所属期刊

推荐期刊

友情链接Friendly link
农业信息研究所Agricultural information institute 农业智能知识服务平台Agricultural intelligent knowledge service platform 农业学术服务平台Agricultural Knowledge Service System 中国知网CNKI 维普网VIP 万方WANFANG DATA
联系我们Contact us

电话Tell：010-82109652-808

邮箱E-mail：agrijournal@caas.cn

服务Service
关于我们About us 意见建议Opinions and suggest
版权说明Copyright
单位：中国农业科学院农业信息研究所 中国农学会 © 2017中国农业科学院农业信息研究所 ICP备案号：京ICP备09089781号-46