大豆种子不同发育时期全长均一化cDNA文库的构建

doi:10.3864/j.issn.0578-1752.2010.03.003

Scientia Agricultura Sinica ›› 2010, Vol. 43 ›› Issue (3): 462-467 .doi: 10.3864/j.issn.0578-1752.2010.03.003

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles Next Articles

Construction of a Normalized Full-Length cDNA Library of Soybean Seed at Different Developmental Stages

LI Chen, YAN Xiao-hong, ZHOU Xin-an, SHA Ai-hua, SHAN Zhi-hui, ZHOU Rong, WEI Wen-hui

（中国农业科学院油料作物研究所基因组学与分子生物学研究室/农业部油料作物生物学重点开放实验室）

Received:2009-06-30 Revised:2009-08-28 Online:2010-02-10 Published:2010-02-10
Contact: WEI Wen-hui

Abstract

Abstract:

【Objective】 The objective of the study is to obtain full-length genes related to seed development and to provide resources for soybean genomics. 【Method】 A normalized cDNA library enriched in full-length sequences was constructed using DSN (duplex-specific nulease)-normalization method combined with SMARTTM(switching mechanism at 5’ end of RNA transcript) technique. 【Result】The titer of unamplified cDNA library was about 6.0×105 cfu/mL. The average cDNA inserts was more than 1.0 kb with a recombination rate of nearly 100%. After large-scale plasmid extraction and sequencing, 36 656 EST sequences were generated from the soybean seed cDNA library and the average length was more than 600 bp. All the EST sequences were assembled and 27 982 unigenes were gotten, which indicated that the cDNA library was a non-redundant library. 【Conclusion】 These results indicate that the normalized full-length cDNA library has been established successfully, which is convenient for further studying the molecular mechanism and gene cloning of seed development.

Key words: soybean, cDNA library, normalization, EST

LI Chen,YAN Xiao-hong,ZHOU Xin-an,SHA Ai-hua,SHAN Zhi-hui,ZHOU Rong,WEI Wen-hui
. Construction of a Normalized Full-Length cDNA Library of Soybean Seed at Different Developmental Stages
[J].Scientia Agricultura Sinica, 2010, 43(3): 462-467 .

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