Construction of a Normalized Full-Length cDNA Library of Sesame Developing Seed by DSN and SMART (TM)

文献类型： 外文期刊

第一作者： Ke Tao

作者： Ke Tao;Dong Cai-hua;Mao Han;Zhao Ying-zhong;Liu Hong-yan;Liu Sheng-yi;Ke Tao

作者机构：

关键词： DSN;full-length library;normalization;oil accumulation;Sesamue indicum;Zhongzhi 14 cDNA library;switching mechanism;SMART (TM)

期刊名称：AGRICULTURAL SCIENCES IN CHINA （ 影响因子：0.82； 五年影响因子：0.997 ）

ISSN： 1671-2927

年卷期： 2011 年 10 卷 7 期

页码：

收录情况： SCI

摘要： Sesame (Sesamue indicum L.) is one of the most important oilseed crops with high oil yield. Here, we described a simple and efficient method for constructing a normalized cDNA library from a high oil content cultivar of sesame Zhongzhi 14, during its oil accumulation stages. It combined switching mechanism at 5'-end of RNA transcript (SMART) technique and duplex-specific nuclease (DSN) normalization methods. Double-stranded cDNAs were synthesized from mRNAs, processed by normalization and Sfi 1 restriction endonuclease, and finally the cDNAs were ligated to pDNR-LIB vector. The ligation mixture was transformed into Escherichia coli DH10B by electroporation. The capacity of the library was 1.0 x 10(6) clones in this library. Gel electrophoresis results indicated the fragments ranged from 700 to 2 000 bp, with the average size of 1800 bp. Random picking clones showed that the recombination rate was 100%. The results showed that the cDNA library constructed successfully was a full-length library with high quality, and could be used to screen the genes related to development of oil synthesis.

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