茉莉酸处理后兴安落叶松针叶cDNA文库的构建及其质量评价  

Construction and Quality Identification of cDNA Library from Larix gmelinii Leaves Induced by Jasmonic Acid

作　　者：金虎[1] 严善春[1] 刘关君[1] 

机构地区：[1]林木遗传育种与生物技术教育部重点实验室(东北林业大学),哈尔滨150040

出　　处：《东北林业大学学报》2010年第6期20-21,56,共3页Journal of Northeast Forestry University

基　　金：国家林业局林业公益性行业科研专项经费资助(200904021)

摘　　要：为研究兴安落叶松(Larix gmelinii)诱导抗虫性的分子机理,以茉莉酸诱导处理后的兴安落叶松针叶为材料,提取叶中总RNA合成cDNA,连接到质粒载体pDNR-LIB上,采用电穿孔法将重组质粒转化到DH5α中。经文库质量鉴定表明:文库库容为5×106,插入片段分布在0.5～2.0kb,平均大小约为1kb,重组率达到95%,表明已成功构建了高质量的茉莉酸诱导处理后兴安落叶松针叶全长cDNA文库。所建文库为进一步开展EST测序分析、抗虫基因克隆及功能基因组学研究奠定了基础。Total RNA was extracted from Larix gmelinii leaves induced by jasmonic acid. cDNA was synthesized and ds-cDNA fragment was ligated into the pDNR-LIB vector. Then the recombinant plasmid was transformed into Escherichia coli DH5α by electroporation. The appraisal of library quality showed that the library had 5×10^6 independent clones,0. 5 ～ 2. 0 kb fragments with an average of about 1 kb in length and a recombination rate of 95% ,indicating that cDNA library of L. gmelinii needles induced by jasmonic acid was constructed successfully. This research lays a base for further study of EST sequencing,full-length cloning of insect-resistant genes,and functional genomics.

关 键 词：茉莉酸 兴安落叶松 CDNA文库 

分 类 号：S791.22[农业科学—林木遗传育种]