烟草突变体<i>T-cldf</i>叶片差异ESTs的克隆与分析

蔡刘体; 郑少清; 叶定勇; 陈雅平; 黄学林

doi:10.3969/j.issn.1002-0861.2008.01.012

烟草突变体T-cldf叶片差异ESTs的克隆与分析

Isolation and Characterization of ESTs Expressed Differently in Leaves of Tobacco Mutant T-cldf

摘要

摘要: 为了获得烟叶生长发育过程中相关的基因信息,以烟草突变体T-cldf叶片为Tester材料,以烟草K346叶片为Driver材料,应用抑制性扣除杂交(Suppression subtractive hybridization,SSH)技术构建了基因差异表达文库,代表在突变体T-cldf叶片中特异表达的cDNA。对随机挑选的部分克隆,经过PCR检测和点杂交筛选的阳性克隆进行序列测定,并与GenBank数据库中的序列进行BLAST比较分析,获得62个表达序列标签(ESTs,已登录GenBank数据库),按照其功能可以分为7类。其中与烟草光合作用相关的ESTs有9个,与结构蛋白相关的有4个,与生理代谢相关的有11个,还有11个基因功能信息未知的ESTs和20个在烟草中首先获得的ESTs。

Abstract: In order to obtain the genic information of tobacco leaf during growth and development, the bank of gene difference expression was constructed with suppression subtractive hybridization (SSH) technology using T-cldf leaves as Tester material and K346 leaves as Driver material, which represented differentially expressed cDNA in T-cldf leaves. The portion randomly selected was cloned and tested by PCR and dot-hybridization screening, the positive clones' sequences were determined and compared with the sequences in GenBank database by BLAST, and 62 expression sequence tags (ESTs, GenBank logged) were obtained. Those ESTs fell into 7 sorts in terms of functions, 9 ESTs related to tobacco photosynthesis, 4 to structural proteins, 11 to physiological metabolism, others included 11 ESTs, the functions of which were unknown, and 20 which were obtained first in tobacco.

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