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学科主题: Plant Sciences
题名: Rapid EST isolation from chromosome IR of rye
作者: Zhou, RN ; Shi, R ; Jiang, SM ; Yin, WB ; Wang, HH ; Chen, YH ; Hu, J ; Wang, RRC ; Zhang, XQ ; Hu, ZM
通讯作者: wawazhoujian@163.com ; rshi@ncsu.edu ; shmjiang@163.com ; wbyin@genetics.ac.cn ; huanhuanbj@hotmail.com ; yhchen@genetics.ac.cn ; jhu@genetics.ac.cn ; richard.wang@ars.usda.gov ; xqzhang@genetics.ac.cn ; zmhu@genetics.ac.cn
刊名: BMC PLANT BIOLOGY
发表日期: 2008
卷: 8, 页:-
收录类别: sci
部门归属: [Zhou, Ruo-Nan; Shi, Rui; Jiang, Shu-Mei; Yin, Wei-Bo; Wang, Huang-Huang; Chen, Yu-Hong; Hu, Jun; Zhang, Xiang-Qi; Hu, Zan-Min] Chinese Acad Sci, Inst Genet & Dev Biol, Beijing 100101, Peoples R China; [Jiang, Shu-Mei] Chinese Acad Sci, S China Sea Inst Oceanol, Guangzhou 510301, Guangdong, Peoples R China; [Wang, Richard R. C.] Utah State Univ, USDA ARS, FRRL, Logan, UT 84322 USA; [Zhou, Ruo-Nan] Chinese Acad Sci, Grad Uni, Beijing 100049, Peoples R China; [Shi, Rui] N Carolina State Univ, Forest Biotechnol Grp, Raleigh, NC 27695 USA
摘要: Background: To obtain important expressed sequence tags (ESTs) located on specific chromosomes is currently difficult. Construction of single-chromosome EST library could be an efficient strategy to isolate important ESTs located on specific chromosomes. In this research we developed a method to rapidly isolate ESTs from chromosome IR of rye by combining the techniques of chromosome microdissection with hybrid specific amplification (HSA). Results: Chromosome IR was isolated by a glass needle and digested with proteinase K (PK). The DNA of chromosome IR was amplified by two rounds of PCR using a degenerated oligonucleotide 6-MW sequence with a Sau3AI digestion site as the primer. The PCR product was digested with Sau3AI and linked with adaptor HSA1, then hybridized with the Sau3AI digested cDNA with adaptor HSA2 of rye leaves with and without salicylic acid (SA) treatment, respectively. The hybridized DNA fragments were recovered by the HSA method and cloned into pMD18-T vector. The cloned inserts were released by PCR using the partial sequences in HSA1 and HSA2 as the primers and then sequenced. Of the 94 ESTs obtained and analyzed, 6 were known sequences located on rye chromosome IR or on homologous group 1 chromosomes of wheat; all of them were highly homologous with ESTs of wheat, barley and/or other plants in Gramineae, some of which were induced by abiotic or biotic stresses. Isolated in this research were 22 ESTs with unknown functions, probably representing some new genes on rye chromosome IR. Conclusion: We developed a new method to rapidly clone chromosome-specific ESTs from chromosome IR of rye. The information reported here should be useful for cloning and investigating the new genes found on chromosome IR.
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WOS记录号: WOS:000255173600001
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内容类型: 期刊论文
URI标识: http://ir.scsio.ac.cn/handle/344004/5368
Appears in Collections:海洋生物_期刊论文

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Zhou, RN; Shi, R; Jiang, SM; Yin, WB; Wang, HH; Chen, YH; Hu, J; Wang, RRC; Zhang, XQ; Hu, ZM.Rapid EST isolation from chromosome IR of rye,BMC PLANT BIOLOGY,2008,8():-
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