茶树基因芯片的研制和初步应用  

Development and Preliminary Application of cDNA Microarray of Tea Plant (Camellia sinensis)

作　　者：赵丽萍[1] 高其康[2] 陈亮[1] 王新超[1] 姚明哲[1] 

机构地区：[1]中国农业科学院茶叶研究所茶树资源遗传改良与分子生物学实验室/农业部茶叶化学工程重点实验室,浙江杭州310008 [2]浙江大学分析测试中心生物大分子研究室,浙江杭州310029

出　　处：《茶叶科学》2006年第3期166-170,共5页Journal of Tea Science

基　　金：浙江省重点科技项目(2004C22033);人事部和教育部留学回国人员科研基金(2005-134;2005-383)

摘　　要：利用EST计划获得的1 680个基因片段,制备了国内外首张茶树cDNA芯片,芯片上每个基因设置一个重复,共含有6 912个点,其中6 720个靶基因,160个阳性对照,32个阴性对照。4×4矩阵点制,共两个区域,每个区域的芯片密度为1 037点/cm2,每张芯片可进行两次杂交。用3个茶多酚含量有差异的品种与芯片进行了杂交,获得了不同品种的基因表达谱及表达差异的基因;选取其中2个与茶叶香气密切相关的基因,采用荧光定量PCR的方法进行了验证,基因表达变化趋势与芯片检测结果一致,验证了cDNA芯片杂交结果的可靠性。该芯片可以进一步应用于许多茶学研究领域,进行基因表达差异的高通量检测。Totally, 1 680 genes obtained in our EST project were selected from the cDNA library of clone Longjing 43 to develop the first cDNA microarray of tea plant. Each gene in the microarray was duplicated. The cDNA microarray contains 6 912 dots, including 6 720 EST, 160 positive controls and 32 negative controls. One microarray was spotted two regions with 4×4 matrixes and it could be hybridized twice. The density of the microarray was 1037 dots per cm^2. Three tea clones with different contents of tea polyphenol were selected to conduct two sets of hybridization experiments with the microarray and the differently expressed ESTs were found. Then, two genes that were closely related to tea aroma were selected for real-time PCR analysis to validate the microarray data and validated the reliability of the cDNA microarray. The newly developed cDNA microarray could be applied in various tea research fields to make the high-throughput detection in the gene expression profiling.

关 键 词：茶树 CDNA芯片 基因表达谱 定量PCR 

分 类 号：S571.1[农业科学—茶叶生产加工]