摘要
以茶树龙井43实生苗新生幼根为材料,应用pBluescript Ⅱ XR cDNA Library Construction Kit,构建了我国第一个茶树幼根cDNA文库。测试结果表明原始文库的滴度为1.85×107pfu/mL,重组率为85.7%,插入片段绝大多数分布在1.0～1.5kb左右。随机挑选5115个克隆进行序列测定,去除空载体和插入片段短于150bp的序列后,获得有效序列4833个,经过序列拼接后共获得3482条表达序列标签(expressed sequence tags,EST),包括901个contigs和2581个singletons。通过与NCBI等非冗余蛋白质数据库进行比对、查询和注释,获得已知功能基因或具推测功能的EST共1376个,代表943个茶树功能基因,421个EST为未知功能基因,1685个EST为新的表达基因。这些数据将为将为我们从分子水平上更好地了解、研究、调控茶树的生理代谢、生长发育和品质等提供极有价值的资源。
The first young roots cDNA library of tea plant was constructed with the young roots of Longjing 43 by using pBluescript II XR cDNA Library Construction Kit. The original library had a high titer of 1.85×10^7 pfu/mL, of which 85.7% clones were recombinant and insert cDNAs were from 1.0 to 1.5 kb. A total of 5 115 clones from the cDNA library of young roots was sequenced randomly and the analysis of these expressed sequence tags （ESTs） were reported herein. A set of 4 833 useful sequences was obtained firstly after excluding clones repeated and shorter than 150 bp. Then clustering and assembly of the 4 833 cDNA sequences generated from the young roots cDNA library resulted in 3 482 unique sequences, including 901 contigs and 2 581 singletons. Among them, 1 376 ESTs with known function or deducible function were identified by Blast X searches against the NCBI non-redundant protein databases, corresponding to more than 943 functional genes in the tea plant. The rest, 421 ESTs were unknown genes, 1 685 ESTs were novel genes partial sequences, respectively. These results acquired from the young roots cDNA library will make it easier to study and understand the physiological metabolism, development and quality of the tea plant on a molecular level.
出处
《分子植物育种》
CAS
CSCD
2008年第5期893-898,共6页
Molecular Plant Breeding
基金
中国863计划(2006AA10Z171)
国家科技支撑计划课题(2006BAD13B06
2006BAD06B01)
浙江省钱江人才计划(2006R10042)资助
关键词
茶树
幼根
CDNA文库
表达序列标签
基因表达谱
Tea plant （Camellia sinensis L. O. Kuntze）, Young roots, eDNA library, Expressed sequence tags （ESTs）, Gene expression profile
